A 32-year-old male had been married for 8 years and attended the Centre for Human Reproduction, Jalandhar, Punjab, India. His childhood development had been normal and uneventful. There was no history of tuberculosis, mumps, injury to the testes and no family history of infertility. Informed consent was obtained before all investigations. He had normal facial but scanty pubic hair, his penile length was normal but the testes were small. The left testis was 17 x 8 mm, whereas the right testis was 21 x 10 mm and each had a volume of 2.0 and 2.5 cc, respectively. Semen analysis showed complete absence of sperm, confirming azoospermia. He had high concentration of serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and low levels of serum testosterone. Serum FSH was 56.18 m IU/mL (normal: 1.4-18.1), LH was 25.83 m IU/mL (normal 1.5-9.3); testosterone total was 179.14 ng/mL (normal: 241-827) and testosterone-free was 2.37 pg/mL (normal 8.80-27.0).

Cytogenetic and Molecular Analysis. Chromosomal preparations were made from phytohemagglutinin-stimulated peripheral lymphocytes using RPMI1640 medium and standard culturing technique with modifications [5]. Chromosomal banding was done by trypsin-Giemsa GTG banding. We examined 100 metaphases for numerical and structural abnormalities and 10 metaphases were karyotyped. All metaphases showed the 46,XX chromosomal complement. Fluorescent in situ hybridization was carried out with an LSI SRY spectrum orange probe according to the manufacturer’s instructions (Vysis UK Ltd., now part of Abbott Molecular, Des Plaines, IL, USA). After hybridization, chromosomes were counterstained with DAPI and viewed by using a triple band-pass filter and Quips Smart Capture Imaging Software (Vysis UK Ltd.). Five hundred metaphase and interphase cells were scored that revealed the presence of the SRY region on cells.