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MOLECULAR AND IN-SITU HYBRIDIZATION DETECTION OF HUMAN PAPILLOMAVIRUS GENOTYPES AMONG EGYPTIAN PATIENTS WITH BLADDER CANCER
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| Author:
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HALA BADAWI, MOHAMMED ABASS, OLFAT HAMAM, MANAL DIAB, MANAL EL SAID, AHMED ISMAIL, AFKAR BADAWY, GEHAN MOSTAFA, SAFIA SAMIR, EHAB EL-DABAA, MOHAMED SABER
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| Abstract:
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Human Papillomavirus (HPV) infection is the most frequent sexually transmitted disease worldwide; more than forty genotypes have shown specific tropism for the male and female genitourinary tract. Aim of this study was to evaluate the sensitivity of reverse slot-blot hybridization (RS-BH) and in-situ hybridization (ISH) for diagnosis and genotyping of HPV infection in cancer bladder patients compared to type-specific PCR (TSPCR) as gold standard.A total of 70 patients with bladder cancer and 30 patients as a control group were conducted in this study. Fresh bladder cancer biopsies were collected from 62 (88.6%) patients with transitional cell carcinoma and 8 (11.4%) with squamous cell carcinoma. HPV-DNA was detected in the biopsies by PCR, RS-BH and ISH. HPV genotyping was performed using TSPCR, RS-BH and ISH. HPV-DNA was detected in 34.3% (24/70) of the tissue specimens by PCR using the general primer set GP5/GP6. HPV genotyping by TSPCR revealed the presence of HPV-16 in 41.7% (10/24) and HPV-18 in 12.5% (3/24). Mixed cases of HPV16-18 genotypes were detected in 12.5% (3/24). ISH results showed that HPV-DNA was detected in 24 out of 70 (34.3%). Nine out of 24 (37.5%) were positive for genotype 16/18. Five specimens (20.8%) were positive for genotype 6/11 and 7 (29.2%) were positive for genotype 31/33. Three (12.5%) mixed cases were detected; 2 (8.3%) were of genotypes 16-18 and 6/11 and one (4.2%) was mixed genotypes 31/33 and 6/11. The results obtained by ISH were similar to TSPCR with absolute sensitivity, specificity, PPV and NPV. Positive HPV genotypes gave signals in 10 out of 24 (41.7%) samples by reverse slot hybridization. HPV genotypes detected were 6, 11, 35, 39, 52 and 56 and were equally distributed. TSPCR and ISH have achieved high sensitivity and specificity in detection of a wide spectrum of HPV types attributed to clinical disease. ISH helped to improve the specificity of the diagnosis and determine the viral genotype which, in turn, helps better define the lesion prognosis
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| Keyword:
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Papillomavirus, Bladder Carcinoma, PCR, genotyping, slot hybridization, in -situ hybridization
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2018.10.04.056
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| Download:
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