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Three cases of rare SRY-negative 46,XX testicular disorder of sexual development with complete masculinization and a review of the literature
J Genet Med 2016;13:78-88
Published online December 31, 2016;
© 2016 Korean Society of Medical Genetics.

Bom Yi Lee1, Shin Young Lee1, Yeon Woo Lee1, Shin Young Kim1, Jin Woo Kim1, Hyun Mee Ryu1,2, Joong Shik Lee3, So Yeon Park1,*, and Ju Tae Seo3,*

1Laboratory of Medical Genetics, Cheil General Hospital and Women’s Healthcare Center, Seoul, Korea,
2Department of Obstetrics and Gynecology, Cheil General Hospital and Women’s Healthcare Center, Dankook University College of Medicine, Seoul, Korea,
3Department of Urology, Cheil General Hospital and Women’s Healthcare Center, Dankook University College of Medicine, Seoul, Korea
Co-corresponding author: Ju Tae Seo, M.D., Ph.D. Department of Urology, Cheil General Hospital and Women’s Healthcare Center, Dankook University College of Medicine, 17 Seoae-ro 1-gil, Jung-gu, Seoul 04619, Korea. Tel: +82-2-2000-7682, Fax: +82-2-2278-4574, E-mail:
So Yeon Park, Ph.D. Laboratory of Medical Genetics, Cheil General Hospital and Women’s Healthcare Center, 17 Seoae-ro 1-gil, Jung-gu, Seoul 04619, Korea. Tel: +82-2-2000-7680, Fax: +82-2-2278-4574, E-mail:
Received May 23, 2016; Revised July 9, 2016; Accepted August 16, 2016.
cc This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.


To identify the clinical characteristics of SRY-negative male patients and genes related to male sex reversal, we performed a retrospective study using cases of 46,XX testicular disorders of sex development with a review of the literature.

Materials and Methods:

SRY-negative cases of 46,XX testicular disorders of sex development referred for cytogenetic analysis from 1983 to 2013 were examined using clinical findings, seminal analyses, basal hormone profiles, conventional cytogenetic analysis and polymerase chain reaction.


Chromosome analysis of cultured peripheral blood cells of 8,386 individuals found 19 cases (0.23%) with 46,XX testicular disorders of sex development. The SRY gene was confirmed to be absent in three of these 19 cases (15.8%).


We report three rare cases of SRY-negative 46,XX testicular disorders of sex development. Genes on autosomes and the X chromosome that may have a role in sex determination were deduced through a literature review. These genes, through differences in gene dosage variation, may have a role in sex reversal in the absence of SRY.

Keywords : Azoospermia, Infertility, SRY genes, 46,XX testicular disorder of sex development

The critical gene for male sex determination, SRY (sex-determining region Y), which is located on chromosome Yp11.3, initiates gonads to differentiate into testes, induces Leydig cells to secrete testosterone, develops Wolffian ducts, and forms male external genitalia. At the same time, Sertoli cells secrete Müllerian inhibiting factor that induces regression of Müllerian ducts that in females would differentiate into the uterus. These processes for sex determination do not occur in females in the absence of SRY. Sex differentiation related genes such as SOX9, FGF9, DAX1, WT1, RSPO1, and SOX10, which are located on either autosomes or the X chromosome, may have a role in gonad development and function. These genes were studied in XX testicular/ovotesticular disorders of sex development (DSDs) in the absence of the SRY gene.

46,XX male sex reversal (also known as testicular DSD) is reported in 1:20,000 to 1:25,000 of newborn males [1], and is categorized using clinical phenotypes or molecular genetic analysis depending on the presence or absence of the SRY gene. Clinical sexual phenotypes of individuals with 46,XX testicular DSD range from complete masculinization to true hermaphrodites (also known as ovotesticular DSD) and are comprised of three groups: 1) XX testicular DSD with normal genitalia of which 85% of cases are normal males with internal and external genitalia, and are usually diagnosed after puberty because of hypogonadism, gynecomastia, and/or infertility [2]; 2) patients with XX testicular DSD with ambiguous genitalia are identified at birth by external genital ambiguities such as micropenises, cryptorchidism, or hypospadias; and 3) patients with XX ovotesticular DSD have internal and external genital ambiguities that are detected at birth or histologically—these patients have either an XX karyotype (60%) or a 46,XX/46,XY or 46,XY karyotypes.

46,XX testicular DSD is also classified genetically based on the presence or absence of the SRY gene [3]. In 90% of cases in which the SRY gene is present, the disorder is the result of an aberrant Y to X chromosomal interchange during meiosis I in paternal gametogenesis. Affected males show a similar phenotype compared to males with Klinefelter syndrome, which is characterized by a 47,XXY karyotype and consists of an active X chromosome, an inactive X chromosome, and the SRY gene. Rarely, the terminal region of the Y chromosome, that includes the SRY gene, is cytogenetically detectable on the X chromosome. For the majority of cases in which the SRY gene is absent, patients with 46,XX testicular DSD have genital ambiguities such as a micropenis, hypospadias, and cryptorchidism. However, completely virilized 46,XX testicular DSD with a SRY deficiency has rarely been reported in the literature. Therefore, other autosomal or X-linked genes may have regulatory roles with SRY in the sex-determination process.

We present three rare cases of SRY-negative 46,XX testicular DSD found in the past 31 years at our center, and our review of the literature on previous cases of 46,XX testicular DSD with a focus on recently reported related genes since the characterization of SRY.

Materials and Methods

1. Clinical findings and cytogenetic analysis

We retrospectively investigated the clinical records of 8,386 males who were referred for cytogenetic analysis from 1983 to 2013 at the Cheil General Hospital, Seoul, Korea. We examined hormonal profiles, histological findings, and semen analysis. For the hormonal profile, we evaluated the levels of serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), 17β-estradiol, testosterone (T), prolactin, and sex hormone-binding globulin.

Conventional cytogenetic analyses were performed according to standard techniques. High-resolution chromosomes of 700-band levels by GTL-banding were prepared using cultured peripheral blood cells. We analyzed 100 metaphases of each patient to exclude potential sex chromosome mosaicism.

Detection of the SRY gene in some patients was confirmed using fluorescence in-situ hybridization (FISH) analysis with the Y-specific probes: CEP Y Sat III Spectrum Green/CEP Y Alpha Spectrum Orange (Vysis Inc., Downers Grove, IL, USA) and the LSI SRY Spectrum Orange/CEP X Spectrum Green (Vysis Inc.) (data not shown).

2. Molecular genetic analysis

Three individuals with 46,XX testicular DSD were further investigated using azoospermia factor (AZF) microdeletion analysis with multiplex polymerase chain reaction (PCR). The oligonucleotides used for multiplex PCR are described in our previous study [4]. Briefly, PCR was performed using the following cycle conditions: an initial denaturation at 95°C for 10 minutes, followed by 35 cycles at 95°C for 10 minutes, 62°C for 90 seconds, and 65°C for 90 seconds with a final extension at 65°C for 10 minutes. Each round of PCR that was performed using patient’s DNA included normal male DNA and normal female DNA as a positive and a negative control, respectively. Amplified products were separated on 3% agarose gel by electrophoresis and visualized using ultraviolet illumination.

3. Ethical considerations

This study was approved by the Ethics Committee of Cheil General Hospital and Women’s Health Care Center (#CGH-IRB-2014-47) and consent was obtained from all patients.


1. Clinical findings and cytogenetic analysis

Of 8,386 male individuals, we identified 1,172 cases with an abnormal karyotype, and 19 cases with 46,XX testicular DSDs. Of these 19 cases, three SRY-negative 46,XX testicular DSDs were confirmed experimentally, and all three cases showed azoospermia with small testes volumes. We found that these 19 patients with 46,XX testicular DSDs have serum concentrations that were partially out of range of normal hormonal profiles. Of those cases of XX testicular DSDs with the SRY gene, we found increased levels of LH, FSH, and T, whereas in cases of XX testicular DSDs without the SRY gene, we found increased levels of LH and FSH with or without decreased levels of T. Patient height ranged from 156 to 172 cm, and we found from our urological assessment that there were no cases of ovotesticular DSD or ambiguous genitalia. With regard to the three cases with 46,XX testicular DSDs without the SRY gene, all three patients showed masculinized external genitalia and small testes with azoospermia. Detailed clinical data is summarized in Table 1.

We analyzed 100 metaphase karyotypes of cultured lymphocytes from peripheral blood of the 19 cases with 46,XX testicular DSDs. All individuals showed a complete 46,XX karyotype. Regardless of the presence or absence of the SRY gene, we found that 0.23% (19/8,386 cases) of the male participants referred for cytogenetic analysis in our study had XX testicular DSDs. Furthermore, we found that 15.8% (3/19 cases) of cases with 46,XX testicular DSDs were SRY-negative or, alternatively, 0.26% (3/1,172 cases) of males with an abnormal karyotype in our cohort of 8,386 participants.

2. Molecular genetic analysis

Of the 16 sequence-tagged site (STS) markers tested, amplification products of 15 STS loci including the SRY gene (sY14) on the Y chromosome were not detected with the exception of ZFX (zinc finger protein, X-linked) located on chromosome Xp21.3, which indicates the complete absence of the Y chromosome (Fig. 1).


Through our study of 8,386 cases and a literature review (summarized in Table 2 [2,5-29]), we found that the major characteristics of male sex reversal for DSD were showed the clinical phenotypes of short stature without a T-dependent pubertal growth spurt and the Y-specific growth gene, azoospermia, internal or external genital abnormalities, gynecomastia, and high levels of FSH and LH, and a low level of T. However, a limitation of our retrospective study was that no gonadal biopsy samples were available for analysis and thus, gonadal mosaicism cannot be ruled out. Reports of familial cases of DSDs in the literature demonstrate that the various clinical phenotypes from ovotesticular DSD to masculinization may be because of incomplete penetrance of one or more genes.

The SRY gene has a critical role in the process of sex differentiation. However, the SOX9, FGF9, and MAP3K1 genes also have roles in the development of testis, the testis-androgen-genital tract to masculinization, anti-Müllerian duct regression, and the duct systems of external genitalia. In addition, RSPO1 and WNT4 function in gonads and have roles in ovary development, the duct system including fallopian tubes, the uterus, and external genitalia without the SRY gene [30,31]. These findings strongly indicate that DSDs with sex reversal involve the balanced regulation of genes in addition to the critical SRY gene, in the cascade of events that lead to development and sex differentiation.

McElreavey et al. [5] proposed the concept of a Z gene product that is a negative regulator of male sex determination. Therefore, homozygosity for loss-of-function and heterozygous mutants of Z alleles induce male sex reversal from SRY-negative complete masculinization to ovotesticular DSD.

Several familial studies have demonstrated that the varying clinical presentation of sexual development such as ovotesticular DSD or XX testicular DSD, may be a result of incomplete penetrance of paternal and maternal inheritance, while dosage-sensitive sex reversal may be a result of autosomal or X-linked genes in the sex-determining cascade [6,7,32].

DAX1 (also known as NROB1), an ‘anti-testis’ factor located on chromosome Xp21 that when duplicated was found to cause XY sex reversal. It has a role in ovarian development and/or in testis formation dependent on dosage-sensitivity during sex determination or sex differentiation pathways in mammals [32,33]. It was found that Dax1 deficiency caused varying degrees of sex reversal in mice [34,35]. In humans, it was found that duplication of the Xp21.3 region containing the DAX1 gene causes testicular regression in the presence of SRY, while the deletion of this region had no effect on male sex determination [36].

Haploinsufficiency of SOX9 mutations have been proposed to cause sex reversal in patients with XY sex reversal, skeletal malformation syndrome, and campomelic dysplasia [37,38]. Huang et al. [8] reported that XX sex reversal was with campomelic dysplasia is caused by duplication of the SOX9 gene, which is located on chromosome 17q24.3. Vernole et al. [9] reported XX testicular DSD with Huriez syndrome and palmoplantar keratoderma, and suggested that a gene related to sex determination that cause Huriez syndrome or tylosis, may be located at the SOX9 locus. Radi et al. [10] reported that male sex reversal with palmoplantar keratoderma/squamous cell carcinoma that was paternally inherited in the same family, would be homozygous for a single gene mutation or contiguous genes. Moreover, increased expression of SOX9 and reduced expression of Ad4BP/SF-1 (also known as NR5A1), DAX1 and anti-Müllerian hormone (AMH, also known as Müllerian inhibiting substance) in SRY-negative XX males, indicates that SOX9 has a key role in sex determination in SRY-negative XX males, and that Ad4BP/SF-1, DAX1 and AMH may contribute to their clinical features [11].

In contrast, Rajender et al. [12] found that there was no evidence of a relationship between SOX9, duplication of 22q, and DAX1 in complete XX testicular DSD without SRY. Temel et al. [13] reported that SOX9 was not duplicated, and no common SOX9 haplotype was shared in nine familial cases with the absence of SRY, and a mutation at the SOX9 locus; the authors proposed that SRY-negative XX testicular DSD may be caused by a monogenic impairment. Maciel-Guerra et al. [14] found no mutations in the SOX9 and DAX1 genes in case with SRY-negative XX maleness (testicular DSD) and monozygotic twins with XX ovotesticular DSD, but showed varying expression of incomplete penetrance of either an autosomal or a X-linked mutation. Seeherunvong et al. [15] investigated 30 subjects without SRY-negative XX testicular or ovotesticular DSD and did not detect any mutations or duplications in the region of chromosome 17q that contains the SOX9 gene using three short tandem repeat (STR) markers.

However, specific critical regions within the SOX9 gene have been recently identified using microarray techniques. Cox et al. [16] found that three family members with 46,XX testicular DSD have a 178-kb microduplication of a gene desert region located 600-kb upstream of SOX9 using SNP-microarray. Vetro et al. [17] found that two brothers who had the same paternal haplotype at the SOX9 region had a 96-kb triplication of a region 500-kb upstream of SOX9 at chromosome 17q24.3 using oligonucleotide array-comparative genomic hybridization. Benko et al. [18] proposed that a 78-kb minimal non-coding region found in a gene desert 517- to 595-kb upstream of the SOX9 promoter included regions suggested by Cox et al. [16] and Vetro et al. [17], and that this region may have one or more gonad-specific SOX9 transcriptional enhancers to induce activation or inactivation of SOX9 gonadal expression in a tissue-specific manner. Xiao et al. [19] narrowed the candidate region to a 74-kb duplication in a 510- to 584-kb region upstream of SOX9 in SRY-negative 46,XX testicular DSD, and proposed based on previous studies, that the candidate region related to gonadal development is a 67-kb region located 584- to 517-kb upstream of SOX9. Recently, there was a report of a SOX9 duplication (~1.45 times) found in a Korean boy with XX testicular DSD [20].

Chiang et al. [21] investigated the FGF9, WT1, NR5A1, and SPRY2 genes in cases with SRY-negative 46,XX testicular DSD and found that FGF9 (located at chromosome 13q12.11) copy number was duplicated compared to that found in normal female controls and was significantly lower that of the normal male controls. Mizuno et al. [22] found that Leydig and Sertoli germ cells have increased expression levels of the Rho-associated, coiled-coil protein kinase 1 (ROCK1) protein, and proposed that testis formation may be regulated by an alternative ROCK1 signaling pathway.

Similar to SOX9, gain of function of the SOX3 (SRY-box 3) gene showed it may be regulated by SRY in sex determination of transgenic mice, and in patients with XX male sex reversal [39]. More recently, Mizuno et al. [23] found copy number gain in the upstream region of the SOX3 gene at chromosome Xq27.1.

A mutation in the RSPO1 (R-spondin 1) gene in patients with 46,XX male sex reversal in an Italian family has demonstrated that sex reversal and palmoplantar hyperkeratosis/squamous cell carcinoma are regulated under via RSPO1 stimulation of keratinocytes and a reduction of β-catenin in the affected keratinocytes [40].

At other autosomal loci, deletion of the short arm of chromosome 11 led to the suggestion that the WT1 (Wilms tumor 1) gene has a role in testis determination [41], while the SF-1 gene has roles in primary adrenal failure and 46,XY gonadal dysgenesis [42]. Aleck et al. [24] reported a partial duplication of 22q13.1 (SOX10) in ovotesticular DSD without SRY. Jiménez et al. [43] proposed ‘vanishing mosaicism’ with 46,XX ovotesticular DSD without SRY; partial deletion of the SRY gene forms a testicular structure, and that an inactivated X chromosome carrying the SRY gene results in the development of ovarian tissue. Mustafa and Mehmet [25] reported an SRY-negative man with complete masculinization and autoimmune thyroiditis, but did not test for any specific genes.

In summary, we report three rare SRY-negative 46,XX testicular DSD cases with complete masculinization and provide a literature review on XX testicular DSD. Although SRY has a master role in sex determination and differentiation, there is a tightly coordinated expression between related genes under SRY for sex development from bipotential gonads. Therefore, the development of maleness in the absence of the SRY gene may be a result of a disruption of this balanced gene expression and gene mutation. Over the past two decades, several genes such as SOX9, SOX3, DAX1, and RSPO1, and chromosomal regions have been reported as potentially critical candidate genes for sex reversal. However, a more comprehensive understanding of other genes that are involved in the underlying networking mechanisms of the sex-determination cascade is necessary.

Fig. 1.

Multiplex-polymerase chain reaction analysis using short tandem repeats for detection of SRY and microdeletion of the azoospermia factor (AZF) region. Samples (p1, p2, and p3) of SRY-negative 46,XX testicular disorders of sex development show complete deletion of the AZFb and AZFc regions. Lanes for the DNA ladder, blank, and female negative controls were loaded and electrophoresed (data not shown).

m, male positive control; p, patient; ZFX, zinc finger protein, X-linked.


Detailed data from 19 cases of 46,XX testicular DSD in 8,386 cases of male individuals referred to cytogenetic analysis from 1983 to 2013

CaseAge (yr)IndicationMarriage (yr)SASRYT (ng/mL)LH (mIU/mL)FSH (mIU/mL)E2 (pg/mL)PRL (ng/mL)SHBG (nmol/L)Free T (pg/mL)Testis R, L (mL)H (cm)/ W (kg)Urological findings
137XXY6AzoNR5, 5Hypoplasia
235 daysAmbiguous genitaliaNR0.1 (0-15)0.8 (2-20)5.6 (2-10)49.5/3.4Bilateral cryptorchidism, hypospadia, pencile type penis, penoplasty; female E, VD, V; (–)R, L
337Infertility4AzoNR3.4 (0-15)9.7 (2-20)22 (2-10)25 (30-120)3, 3Penis (5 cm) E, VD, V; (–)R, L
436Infertility3AzoNR1.9 (0.2-0.8)18 (1.8-13.4)42 (2-12)13 (30-120)8, 8165/59penis (8 cm) E, VD, V; (–)R, L
528Infertility3Azo(+)3.6 (0.2-0.8)22 (8.0-13.4)64 (2-12)7.8 (0-15)4, 4165/55Penis (7 cm) E, VD, V; (–)R, L
631Infertility9AzoNR2.2 (0.2-0.8)14 (1.8-13.4)42 (2-12)3, 3Penis (4 cm), tubular sclerosis & hyalinization, focal increased number of Leydig cells
730InfertilityNR5.1 (0.2-0.8)4.5 (1.8-13.4)16 (2-12)12 (30-120)10, 10E, VD, V; (–)R, L
8335AzoNR4, 4/65Germ cell aplasia
937Infertility4AzoNR4.7 (0.2-0.8)17 (1.8-13.4)48 (2-12)19 (10-73)18 (8.8-27)2, 2162/65Penis (6 cm), E, VD, V; (–)R, L
1033Infertility3AzoNR1.95 (2.5-8.8)17.2 (0-12)22.3 (0-15)3, 3174/73E, VD, V; (–)R, L
1138Infertility4Azo(+)3.5 (0.2-0.8)18 (1.8-13.4)45 (2-12)4.3 (0-15)3, 3160/62Leydig cell hyperplasia, AZF gene microdeletion E, VD, V; (–)R, L
1228Infertility3AzoNR1.8 (0.2-0.8)13 (1.8-13.4)29 (2-12)12, 12Germ cell aplasia, R. testicular atrophy, bilateral retractile testis, bilateral orchiopexy (undescended) E, VD, V; (–)R, L
1336Infertility7Azo(+)2.1 (0.2-0.8)13 (1.8-13.4)35 (2-12)35 (10-73)13 (8.8-27)3, 3173/76Penis (9 cm), AZF gene microdeletion
1429Infertility1Azo(+)2.78 (1.3-8.1)9.2 (1.5-9.2)44 (1-14)11 (15-80)3.8 (0-15)25 (10-73)5, 5173/80Germ cell aplasia (60-70%) atrophy (30-40%) E, VD, V; (–)R, L
1529InfertilityAzo(+)1.3 (1.3-8.1)5.9 (1.5-9.2)34 (1-14)8 (15-80)8.8 (2.7-19.7)11 (10-73)2, 2163/72E, VD, V; (–)R, L
1641Azo4.11Azo(+)1.27 (1.3-8.1)30.7 (1.7-8.6)48.8 (1.5-12.4)9.9 (7.4-42.6)7.6 (4.0-15.2)39.8 (10-73)2.09 (8.8-27)2, 2163/60E, VD, V; (–)R, L
1737Infertility6.11Azo(–)0.58 (1.3-8.1)8.4 (1.7-8.6)5.3 (1.5-12.4)144.7 (7.4-42.6)14.4 (4.0-15.2)79 (10-73)2, -165/56Male hormonal treatment E, VD, V; (–)R, L
1842Known XX5Azo(–)1.62 (1.3-8.1)13.3 (1.7-8.6)45.1 (1.5-12.4)8.1 (4.0-15.2)38.3 (10-73)3.2 (8.8-27)2, 2156/45Male hormonal treatment E, VD, V; (–)R, L
1936Azo6Azo(–)0.47 (1.3-8.1)19.1 (1.7-8.6)27 (1.5-12.4)17 (7.4-42.6)2.9 (4.0-15.2)25.4 (10-73)0.8 (8.8-27)2, 2172/78Male hormonal treatment E, VD, V; (–)R, L

DSD, disorders of sex development; SA, semen analysis; LH, luteinizing hormone; FSH, follicle-stimulating hormone; E2, 17β-estradiol; PRL, prolactin; SHBG, sex hormone-binding globulin; T, testosterone; R, right; L, left; H, height; W, weight; Azo, azoospermia; NR, not reported; E, epididymis; VD, vas deferens; V, varicocele; (+), present; (–), absent; AZF, azoospermic factor.

Summary of previously reported SRY-negative males with 46,XX testicular DSD or ovotesticular DSD

CaseYear [ref]Age (yr)PhenotypeHormoneDiagnosisInheritanceTested/proposed candidate gene(s)
11993 [5]Normal male without ambiguityXX TDfam, sib/Z gene theory, Z-/Z-, Z+/Z-, Z+/Z+; recessive mutations, wild type Z product, a negative regulator of male sex determination that is functional in wild-type female
2Hypospadias, micropenis, hyperclitoridyXX TDfam, sib
3Internal & external genital ambiguityOTDfam, sib
41993 [6]>24Normal penis, small testes, sister & mat cousin sister with OTDFSH↑XX TDfam, matPABY, SRY, ZFY/ autosomal dominant mutation or X-linked dominant gene downstream from SRY, incomplete penetrance
51997 [2]286-cm penis, both gonads (3.78, 4.13 mL), sparse facial, body, & axillary hair, female pubic hair (Tanner stage 4), gynecomastia, hypergonadotropic hypogonadism, height 156 cmXX TDfam, sibPABY, SRY, ZFY/ a homozygous loss-of-function mutation in a recessive, autosomal or X-linked gene, resulting in activation of the male sex-determining pathway
6267-cm penis, both gonads (3.3, 4.12 mL), abundant facial, body, & axillary hair, female pubic hair (Tanner stage 4), height 162 cmXX TDfam, sib
71998 [7]4 wkSmall penis, hypospadias, a bifid scrotum, nonpalpable testes, intra-abdominal gonad (R), gonadal remnant (L) small midline uterus, vaginaT↓OTDfam, mat/Autosomal or X-linked mutation, the different phenotypic effects arise because of variable penetrance
812No pubertal signs, height in the 10th centileXX TDfam, mat
91999 [8]Infant1.2-cm penis, scrotal hypospadias, the opening urethral meatus, bifid scrotum, palpable gonads, uterus (–), 46,XX,dup(17) (q23.1q24.3)/46,XX,dnXX TD/An extra dose of SOX9 is sufficient to initiate testis differentiation in SRY(–)
101999 [24]InfantSmall penis, scrotal sack (R), pigmented scrotal tissue (L), small vagina, uterus, & ovary, streak gonad (L), a small testis-like structure with VD & E (R), 46,XX,rec(22)dup(22q) inv(22)(p13q13.1)matT↑OTDPartial dup of chr 22/ genes on chr 22 that are involved in sex determination
112000 [9]42Sparse facial, body, axillary hair & male pubic hair, hypospadias, cryptorchidism, normal heightFSH↑ T↓XX TD/Autosomal or X-linked sex-determining genes, both carriers for a recessive mutated allele in the SOX9 locus, Huriez syndrome, sclerotylosis
12-172004 [26]6 CasesXX TDOver expression of the SOX10 gene at 22q13 in a sex reversal case; no evidence in 13 additional subjects with SRY(–) 46,XX sex reversal for microduplication of 22q (SOX10) /a gene on 22q that can trigger testis determination in the absence of SRY
18-247 CasesOTD
252005 [10]Hypospadias & hypogenitalism, gynecomastia, PPK/SSCXX TDfam, sibLinkage analysis of 15 loci for PPK and 9 loci for sex determination /differentiation a single mutation/ possibly affecting contiguous genes may underlie both sex reversal & PPK/SCC. The SOX9 gene is very close to the locus for “tylosis”.
26Hypospadias & hypogenitalism, PPKXX TD
27Hypospadias & hypogenitalism, two epididymal cysts (4.5 mm), gynecomastia, nodular hyperplasia of Leydig cells. PPK/SCCXX TD
282006 [12]34Testes (4.8, 5.1 mL), normal axillary & pubic hair, infertility, height 156 cmFSH↑XX TDSOX9, DAX1, 22q; no mutation, no dup in SOX9- 17q (6 STRs), 22q (3 STRs), no deletion in DAX1-X (53 STRs)/ a loss of function mutation in a ‘gene’ downstream to SRY in the male determining pathway
292007 [13]154-5-cm penis, severe chordee, penoscrotal hypospadias, female pubic hair, blind ending vagina, palpable gonadsXX TDfama mutation at a sex-determining locus other than SRY & SOX9 as the cause for the XX sex reversal trait in this family, no a common SOX9 haplotype identified among family members
30134-5-cm penis, labiascrotal fusion, scrotal hypospadias, hypoplastic hyposcrotumOTD
3153-4-cm penis, severe chordee, bifid scrotum, nonpalpable R. gonad, penoscrotal hypospadias. vaginal orifice (3 cm), penoscrotal transpositionOTD
32153-4-cm penis, severe chordee, perineal hypospadias, nonpalpable gonads, female pubic hairOTD
3333Normal penis, infertility, proximal glandular hypospadias, palpable gonadsFSH↑XX TD
3410Normal penis & meatus, palpable scrotum, female pubic hairNormal FSH, LH, TXX TD
3526Normal penis, megameatus, infertility, palpable gonadsFSH↑XX TD
3624Normal penis & meatus, palpable gonadsFSH↑XX TD
3719Normal penis size, distal glandular hypospadias, palpable gonadsFSH↑XX TD
382008 [14]Infant1-cm penis, chordee, penoscrotal hypospadias, palpable gonadsNormal FSH, LH, TXX TDfam, twinSOX9, DAX1; no mutation
39Infant0.5-cm penis, perineal hypospadias, gonad (R, nonpalpable; L, small)FSH↑OTDfam, twin
402008 [11]235-cm penis, hypospadias, testes (5.1, 0 mL), height 157 cmFSH↑, T↓OTDSOX9, DAX1, Ad4BP/SF-1, WT1, GATA4, MIS. SOX9 expression↑ and expression↓ of Ad4BP/SF-1, DAX1 & MIS /lesions affecting SOX9 expression are the key factor in sex determination in SRY(–) XX males, the decreased expression of Ad4BP/SF-1, DAX-1 & MIS contribute to their clinical features
41283.6-cm penis, hypospadias, testes (3.3, 3.2 mL), height 150 cmFSH↑, T↓OTD
42213.6-cm penis, hypospadias, testes (4.2, 3.8 mL), height 156 cmFSH↑, T↓OTD
435Chordee, hypospadias (–)OTD
44203.2-cm penis, hypospadias, testes (4.2, 3.1 mL)OTD
452010 [27]29Azo, testes (3, 5 mL), Leydig cell hyperplasia, height 165 cmFSH↑, T↓XX TD/25 cases review, a number of unknown genes downstream participate in sex determination
462010 [25]30Azo, small testes & scrotum, normal axillary & pubic hair, gynecomastia, hypo-thyroidism, height 170 cmFSH↑, TSH↑, T↓XX TD/Chronic autoimmune thyroiditis
472011 [16]AdultAzo, normal virilization, Leydig & Sertoli cells↓XX TDfam, pat178-kb dup. 600-kb upstream of SOX9, gene desert region/ autosomal dominant sex-limited inheritance
48AdultXX TDfam, pat
49AdultAzo, normal virilizationXX TDfam, pat
502011 [17]47Azo, hypotrophic testes, germinal cell aplasia, mild gynecomastiaFSH↑, T↓XX TDfam, pat96-kb triplication 500-kb upstream of SOX9 / cis-acting regulatory elements located within the smaller XX-sex reversal critical region; dup. increase SOX9 expression driving testicular differentiation in SRY(–)
5146Azo, hypotrophic testes, germinal cell aplasia, mild gynecomastiaFSH↑, T↓XX TDfam, pat
522011 [18]Infant1.3-cm penis, hypospadias, bifid scrotum, male external genitalia, palpable gonads, bilateral fallopian tubesFSH↑, T↑OTDSOX9; minimum 78-kb dup located in gene desert region 517-595-kb upstream of the SOX9 promoter/gonad specific SOX9 transcriptional enhancer(s), the gain or loss of this region may act as a sex-determination switch in a tissue specific manner
53InfantPerineal hypospadias, asymmetric scrotum, L. scrotal with an ovarian remnant & fallopian tubes, surgery of a vagina & uterusOTDfam, mat
54InfantPerineal hypospadias, 2.5-cm curved penis, hypoplastic & asymmetric scrotum, R. palpable gonad, vaginal pouch & uterusFSH↑OTDfam, pat
55-772012 [15]The 23 subjects had range in the extent range of masculinization of the external genitaliaXX TDSOX9-17q23.1-q24.3; no dup/ dup of SOX9 is not a common cause, microduplication or rearrangement of the SOX3 locus is a more common cause of 46,XX testicular & 46,XX ovotesticular DSD
78-84Small penis, penoscrotal/perineal hypospadias, bilateral ovotestesOTD
852013 [21]52Azo, small testes, glandular hypospadia, virilization, height 160.3 cmFSH↑, T↓XX TDAmong FGF9, WT1, NR5A1, SPRY2, dup of FGF9 & increase of SPRY2 gene copy number/ may hinder WNT4 expression and delaying of ovarian development in XX testicular DSD
862013 [28]403.6-cm penis, Azo, testes (2, 2.5 mL), male pubic hair (Tanner stage 4), hypergonadotropic hypogonadismNormal T, E2, FSH↓,XX TD/Review, management guidance
872013 [19]27Azo, small testes, correction of congenital hypospadiasFSH↑, T↓XX TDDAX1, SOX9, RSPO1; ~74 kb dup upstream of SOX9 without mutation/ ~67 kb critical region of may lead to SOX9 overexpression, causing female-to-male sex reversal
2013 [22]Genital ambiguity, hypospadias, bilateral cryptorchidism (two cases of Kojima et al. [11])11 ↑expressed genes (ROCK1, PQBP1, UCP2, OR13G1, ZNFX1, MPHOSPH8, GNRHR2, YIPF6, HSP90AB2P, KIF27, and YIF1B) & 7 ↓expression genes (EEF1A1, FTH1, UBB, RPS25, RPL7a, RPL6, and RPL32); ↑expression of ROCK1 in XX male, ROCK1 phosphorylates & activates SOX9 in Sertoli cells/ Testes formation by an alternative signaling pathway & ROCK1
2014 [23]Four cases of Kojima et al. [11] and Mizuno et al. [22]High SOX3 gene expression (Xq27.1) leads to testicular differentiation despite SRY(–)
882014 [20]4No gross anomalies, small testes without ambiguity, height 98.2 cm (10th-25th)n LH, FSH, E2, 17-OHP, hCG,XX TDCopy number dup of SOX9 (1.44-1.45:1)
892014 [29]14Congenital scrotal type hypospadias, gynecomastia IA, Azo, small penis and testes, height 155 cm (age mean 165.9±7.21 yr )nT, E2, PRL, FSH↑,XX TDNo mutation of DAX1, SOX9, SOX3, SOX10, ROCK1, and DMRT, and no copy number variation in whole genome
902015 [current study]37Azo, testes (2,–mL), [E, VD, V R(–)], height 165 cmE2↑, SHBG, T↓XX TDThe current study
9142Azo, testes (2, 2 mL), [E, VD, V R(–), L(–)], height 156 cmFSH↑, T↓XX TD
9236Azo, testes (2, 2 mL), [E, VD, V R(–), L(–)], height 172 cmFSH↑, T↓, PRL↓XX TD

DSD, disorders of sex development; ref, reference number; TD, testicular DSD; OTD, ovotesticular DSD; fam, familial; sib, sibling; FSH, follicle stimulating hormone; LH, luteinizing hormone; R, right; L, left; T, testosterone; chr, chromosome; STRs, short tandem repeats; PPK, palmoplantar keratoderma; TSH, thyroid-stimulating hormone; mat, maternal; SCC, squamous cell carcinoma; Azo, azoospermia; dup, duplication; hCG, human chorionic gonadotropin; E2, 17β-estradiol; 17-OHP, 17-hydroxyprogesterone; pat, paternal; PRL, prolactin; SHBG, sex hormone-binding globulin; E, epididymis; V, varicocele; VD, vas deferens; (+), present; (–), absent; (↓), decreased; (↑), increased.

  1. de la Chapelle A. Analytic review: nature and origin of males with XX sex chromosomes. Am J Hum Genet 1972;24:71-105.
    Pubmed KoreaMed
  2. Zenteno JC, López M, Vera C, Méndez JP, and Kofman-Alfaro S. Two SRY-negative XX male brothers without genital ambiguity. Hum Genet 1997;100:606-10.
    Pubmed CrossRef
  3. Ferguson-Smith MA, Cooke A, Affara NA, Boyd E, and Tolmie JL. Genotype-phenotype correlations in XX males and their bearing on current theories of sex determination. Hum Genet 1990;84:198-202.
    Pubmed CrossRef
  4. Lee BY, Kim SY, Park JY, Choi EY, Kim DJ, and Kim JW et al. Unusual maternal uniparental isodisomic x chromosome mosaicism with asymmetric y chromosomal rearrangement. Cytogenet Genome Res 2014;142:79-86.
    Pubmed CrossRef
  5. McElreavey K, Vilain E, Abbas N, Herskowitz I, and Fellous M. A regulatory cascade hypothesis for mammalian sex determination: SRY represses a negative regulator of male development. Proc Natl Acad Sci U S A 1993;90:3368-72.
    Pubmed KoreaMed CrossRef
  6. Kuhnle U, Schwarz HP, Löhrs U, Stengel-Ruthkowski S, Cleve H, and Braun A. Familial true hermaphroditism: paternal and maternal transmission of true hermaphroditism (46,XX) and XX maleness in the absence of Y-chromosomal sequences. Hum Genet 1993;92:571-6.
    Pubmed CrossRef
  7. Slaney SF, Chalmers IJ, Affara NA, and Chitty LS. An autosomal or X linked mutation results in true hermaphrodites and 46,XX males in the same family. J Med Genet 1998;35:17-22.
    Pubmed KoreaMed CrossRef
  8. Huang B, Wang S, Ning Y, Lamb AN, and Bartley J. Autosomal XX sex reversal caused by duplication of SOX9. Am J Med Genet 1999;87:349-53.
  9. Vernole P, Terrinoni A, Didona B, De Laurenzi V, Rossi P, and Melino G et al. An SRY-negative XX male with Huriez syndrome. Clin Genet 2000;57:61-6.
    Pubmed CrossRef
  10. Radi O, Parma P, Imbeaud S, Nasca MR, Uccellatore F, and Maraschio P et al. XX sex reversal, palmoplantar keratoderma, and predisposition to squamous cell carcinoma: genetic analysis in one family. Am J Med Genet A 2005;138A:241-6.
    Pubmed CrossRef
  11. Kojima Y, Hayashi Y, Mizuno K, Sasaki S, Fukui Y, and Koopman P et al. Up-regulation of SOX9 in human sex-determining region on the Y chromosome (SRY)-negative XX males. Clin Endocrinol (Oxf) 2008;68:791-9.
    Pubmed CrossRef
  12. Rajender S, Rajani V, Gupta NJ, Chakravarty B, Singh L, and Thangaraj K. SRY-negative 46,XX male with normal genitals, complete masculinization and infertility. Mol Hum Reprod 2006;12:341-6.
    Pubmed CrossRef
  13. Temel SG, Gulten T, Yakut T, Saglam H, Kilic N, and Bausch E et al. Extended pedigree with multiple cases of XX sex reversal in the absence of SRY and of a mutation at the SOX9 locus. Sex Dev 2007;1:24-34.
    Pubmed CrossRef
  14. Maciel-Guerra AT, de Mello MP, Coeli FB, Ribeiro ML, Miranda ML, and Marques-de-Faria AP et al. XX Maleness and XX true hermaphroditism in SRY-negative monozygotic twins: additional evidence for a common origin. J Clin Endocrinol Metab 2008;93:339-43.
    Pubmed CrossRef
  15. Seeherunvong T, Ukarapong S, McElreavey K, Berkovitz GD, and Perera EM. Duplication of SOX9 is not a common cause of 46,XX testicular or 46,XX ovotesticular DSD. J Pediatr Endocrinol Metab 2012;25:121-3.
    Pubmed CrossRef
  16. Cox JJ, Willatt L, Homfray T, and Woods CG. A SOX9 duplication and familial 46,XX developmental testicular disorder. N Engl J Med 2011;364:91-3.
    Pubmed CrossRef
  17. Vetro A, Ciccone R, Giorda R, Patricelli MG, Della Mina E, and Forlino A et al. XX males SRY negative: a confirmed cause of infertility. J Med Genet 2011;48:710-2.
    Pubmed KoreaMed CrossRef
  18. Benko S, Gordon CT, Mallet D, Sreenivasan R, Thauvin-Robinet C, and Brendehaug A et al. Disruption of a long distance regulatory region upstream of SOX9 in isolated disorders of sex development. J Med Genet 2011;48:825-30.
    Pubmed CrossRef
  19. Xiao B, Ji X, Xing Y, Chen YW, and Tao J. A rare case of 46, XX SRY-negative male with approximately 74-kb duplication in a region upstream of SOX9. Eur J Med Genet 2013;56:695-8.
    Pubmed CrossRef
  20. Lee GM, Ko JM, Shin CH, and Yang SW. A Korean boy with 46,XX testicular disorder of sex development caused by SOX9 duplication. Ann Pediatr Endocrinol Metab 2014;19:108-12.
    Pubmed KoreaMed CrossRef
  21. Chiang HS, Wu YN, Wu CC, and Hwang JL. Cytogenic and molecular analyses of 46,XX male syndrome with clinical comparison to other groups with testicular azoospermia of genetic origin. J Formos Med Assoc 2013;112:72-8.
    Pubmed CrossRef
  22. Mizuno K, Kojima Y, Kamisawa H, Moritoki Y, Nishio H, and Kohri K et al. Gene expression profile during testicular development in patients with SRY-negative 46,XX testicular disorder of sex development. Urology 2013;82:Array-7.
    Pubmed CrossRef
  23. Mizuno K, Kojima Y, Kamisawa H, Moritoki Y, Nishio H, and Nakane A et al. Elucidation of distinctive genomic DNA structures in patients with 46,XX testicular disorders of sex development using genome wide analyses. J Urol 2014;192:535-41.
    Pubmed CrossRef
  24. Aleck KA, Argueso L, Stone J, Hackel JG, and Erickson RP. True hermaphroditism with partial duplication of chromosome 22 and without SRY. Am J Med Genet 1999;85:2-4.
  25. Mustafa O, and Mehmet E. A 46, XX SRY - negative man with infertility, and co-existing with chronic autoimmune thyroiditis. Gynecol Endocrinol 2010;26:413-5.
    Pubmed CrossRef
  26. Seeherunvong T, Perera EM, Bao Y, Benke PJ, Benigno A, and Donahue RP et al. 46,XX sex reversal with partial duplication of chromosome arm 22q. Am J Med Genet A 2004;127A:149-51.
    Pubmed CrossRef
  27. Kim JW, Bak CW, Chin MU, Cha DH, Yoon TK, and Shim SH. SRY-negative 46,XX infertile male with Leydig cell hyperplasia: clinical, cytogenetic, and molecular analysis and review of the literature. Fertil Steril 2010;94:Array-9.
    Pubmed CrossRef
  28. Ryan NA, and Akbar S. A case report of an incidental finding of a 46,XX, SRY-negative male with masculine phenotype during standard fertility workup with review of the literature and proposed immediate and long-term management guidance. Fertil Steril 2013;99:1273-6.
    Pubmed CrossRef
  29. Li TF, Wu QY, Zhang C, Li WW, Zhou Q, and Jiang WJ et al. 46,XX testicular disorder of sexual development with SRY-negative caused by some unidentified mechanisms: a case report and review of the literature. BMC Urol 2014;14:104.
    Pubmed KoreaMed CrossRef
  30. Hughes IA. Disorders of sex development: a new definition and classification. Best Pract Res Clin Endocrinol Metab 2008;22:119-34.
    Pubmed CrossRef
  31. Vinci G, Brauner R, Tar A, Rouba H, Sheth J, and Sheth F et al. Mutations in the TSPYL1 gene associated with 46,XY disorder of sex development and male infertility. Fertil Steril 2009;92:1347-50.
    Pubmed CrossRef
  32. Bardoni B, Zanaria E, Guioli S, Floridia G, Worley KC, and Tonini G et al. A dosage sensitive locus at chromosome Xp21 is involved in male to female sex reversal. Nat Genet 1994;7:497-501.
    Pubmed CrossRef
  33. Zanaria E, Muscatelli F, Bardoni B, Strom TM, Guioli S, and Guo W et al. An unusual member of the nuclear hormone receptor superfamily responsible for X-linked adrenal hypoplasia congenita. Nature 1994;372:635-41.
    Pubmed CrossRef
  34. Bouma GJ, Albrecht KH, Washburn LL, Recknagel AK, Churchill GA, and Eicher EM. Gonadal sex reversal in mutant Dax1 XY mice: a failure to upregulate Sox9 in pre-Sertoli cells. Development 2005;132:3045-54.
    Pubmed CrossRef
  35. Park SY, Lee EJ, Emge D, Jahn CL, and Jameson JL. A phenotypic spectrum of sexual development in Dax1 (Nr0b1)-deficient mice: consequence of the C57BL/6J strain on sex determination. Biol Reprod 2008;79:1038-45.
    Pubmed KoreaMed CrossRef
  36. Sarafoglou K, and Ostrer H. Clinical review 111: familial sex reversal: a review. J Clin Endocrinol Metab 2000;85:483-93.
    Pubmed CrossRef
  37. Foster JW, Dominguez-Steglich MA, Guioli S, Kwok C, Weller PA, and Stevanovic´ M et al. Campomelic dysplasia and autosomal sex reversal caused by mutations in an SRY-related gene. Nature 1994;372:525-30.
    Pubmed CrossRef
  38. Wagner T, Wirth J, Meyer J, Zabel B, Held M, and Zimmer J et al. Autosomal sex reversal and campomelic dysplasia are caused by mutations in and around the SRY-related gene SOX9. Cell 1994;79:1111-20.
  39. Sutton E, Hughes J, White S, Sekido R, Tan J, and Arboleda V et al. Identification of SOX3 as an XX male sex reversal gene in mice and humans. J Clin Invest 2011;121:328-41.
    Pubmed KoreaMed CrossRef
  40. Parma P, Radi O, Vidal V, Chaboissier MC, Dellambra E, and Valentini S et al. R-spondin1 is essential in sex determination, skin differentiation and malignancy. Nat Genet 2006;38:1304-9.
    Pubmed CrossRef
  41. Call KM, Glaser T, Ito CY, Buckler AJ, Pelletier J, and Haber DA et al. Isolation and characterization of a zinc finger polypeptide gene at the human chromosome 11 Wilms’tumor locus. Cell 1990;60:509-20.
  42. Achermann JC, Ito M, Ito M, Hindmarsh PC, and Jameson JL. A mutation in the gene encoding steroidogenic factor-1 causes XY sex reversal and adrenal failure in humans. Nat Genet 1999;22:125-6.
    Pubmed CrossRef
  43. Jiménez AL, Kofman-Alfaro S, Berumen J, Hernández E, Canto P, and Méndez JP et al. Partially deleted SRY gene confined to testicular tissue in a 46,XX true hermaphrodite without SRY in leukocytic DNA. Am J Med Genet 2000;93:417-20.

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