Transplant Recipients and Anal Neoplasia Study: Design, Methods, and Participant Characteristics of a Prevalence Study

Kidney recipients have anal cancer rates 3 times higher than the general population in Australia and New Zealand. High-risk human papillomavirus (HPV) genotypes are implicated in the majority of anal cancers. Establishing the epidemiology of anal HPV infection and precursors of anal cancer in transplant recipient populations is 1 consideration in any potential screening program. The Transplant and Anal Neoplasia Study is a cross-sectional study of the prevalence of anal cytological abnormalities and HPV deoxyribonucleic acid in kidney transplant recipients, as well as evaluating the acceptability of an anal cancer screening intervention. The study aims to recruit 100 kidney transplant recipients, older than 18 years, in Australia. Transplant recipients at- tending for a protocol biopsy at 3 and 12 months and annually posttransplant are approached to participate. Participants undergo an anal swab, which is then analyzed using liquid-based cytological examination and tested for the detection of 37 anogenital HPV deoxyribonucleic acid genotypes. Participants also complete a demographic and behavioral questionnaire that covers sexual be- havior, history of anal symptoms, and possible anal cancer risk factors. Associations will be tested using multiple regression anal- ysis. Recruitment for the study began in 2015 and is ongoing. To date, 96 (77%) of 125 kidney transplant recipients approached have consented to the study. The mean age is 48 (median, 47 y; range, 20–76 y), 59% are male, and Northwest European (58%) represented the largest ethnic group. No participants self-identified as Aboriginal or Torres Strait Islander. High consent rates and positive qualitative results suggest that a larger screening program may be well received by kidney transplant recipients, with in- creased resources and some modification to the timing of approach. Further results of the study will inform the possible implemen- tation of a larger screening trial for prevention of anal cancers in kidney and other solid organ transplant recipients

Development of a rapid colorimetric multiplex PCR-reverse line blot for the detection and typing of 14 Chlamydia trachomatis genovars

Purpose. Chlamydiatrachomatis is responsible for trachoma-associated blindness as well as the most common sexually transmitted bacterial infection worldwide, although the genovars for the former are typically A-C, whilst for the latter they are D-K and for the uncommon infection lymphogranuloma venereum they are L1-3. Nucleotide variations within the ompA gene facilitate the identification of C. trachomatis genovars. This study describes a colorimetric multiplex PCR/RLB typing assay (mPCR-RLB) directed to the VD2 region of the ompA gene for general C. trachomatis positivity and the identification of 14 individual C. trachomatis genovars. Methodology. The assay was validated by analysing 40 blinded samples that included reference strains of C. trachomatis genovars and other non-chlamydial micro-organisms that had been analysed previously using quantitative PCR (qPCR). Ninety clinical samples that had previously been found to be C. trachomatis-positive by qPCR were also evaluated using the mPCR-RLB assay. Results. The mPCR-RLB assay showed 100% agreement with the qPCR in the detection of C. trachomatis reference strains and no cross-reaction of non-chlamydial micro-organisms was observed. In the analysis of the chlamydial clinical samples, 97.8% were C. trachomatis-positive by mPCR/RLB assay and there was a 96.6% concordance with the qPCR at the group identification level and a 92.2% concordance at the genovar level. Conclusion. The mPCR-RLB assay is a rapid and sensitive methodology for the identification of C. trachomatis genovars associated with urogenital infections, trachoma or lymphogranuloma venereum diseases that can be implemented in clinical settings, helping to identify reinfections and treatment failures and establish the appropriate treatment course

HPV genotype-specific concordance between EuroArray HPV, Anyplex II HPV28 and Linear Array HPV Genotyping test in Australian cervical samples

Purpose: To compare human papillomavirus genotype-specific performance of two genotyping assays, Anyplex II HPV28 (Seegene) and EuroArray HPV (EuroImmun), with Linear Array HPV (Roche). Methods: DNA extracted from clinican-collected cervical brush specimens in PreservCyt medium (Hologic), from 403 women undergoing management for detected cytological abnormalities, was tested on the three assays. Genotype-specific agreement were assessed by Cohen's kappa statistic and Fisher's z-test of significance between proportions. Results: Agreement between Linear Array and the other 2 assays was substantial to almost perfect (κ = 0.60 â 1.00) for most genotypes, and was almost perfect (κ = 0.81 â 0.98) for almost all high-risk genotypes. Linear Array overall detected most genotypes more frequently, however this was only statistically significant for HPV51 (EuroArray; p = 0.0497), HPV52 (Anyplex II; p = 0.039) and HPV61 (Anyplex II; p=0.047). EuroArray detected signficantly more HPV26 (p = 0.002) and Anyplex II detected more HPV42 (p = 0.035) than Linear Array. Each assay performed differently for HPV68 detection: EuroArray and LA were in moderate to substantial agreement with Anyplex II (κ = 0.46 and 0.62, respectively), but were in poor disagreement with each other (κ = â0.01). Conclusions: EuroArray and Anyplex II had similar sensitivity to Linear Array for most high-risk genotypes, with slightly lower sensitivity for HPV 51 or 52. Keywords: Human papillomavirus, Genotyping, Linear Array, Anyplex II, EuroArray, Cervi

Accuracy of Self-reported Human Papillomavirus Vaccination Status Among Gay and Bisexual Adolescent Males: Cross-sectional Study

BackgroundMen who have sex with men are a risk group for anal human papillomavirus (HPV) and anal cancer. Australia introduced a universal school-based HPV vaccination program in 2013. Self-reported HPV vaccination status has been widely used in clinical and research settings, but its accuracy is understudied. ObjectiveWe aimed to examine the accuracy of self-reported HPV vaccination status among gay and bisexual adolescent males. MethodsWe included 192 gay and bisexual males aged 16-20 years from the Human Papillomavirus in Young People Epidemiological Research 2 (HYPER2) study in Melbourne, Australia. All participants had been eligible for the universal school-based HPV vaccination program implemented in 2013 and were asked to self-report their HPV vaccination status. Written informed consent was obtained to verify their HPV vaccination status using records at the National HPV Vaccination Program Register and the Australian Immunisation Register. We calculated the sensitivity, specificity, positive predictive value, and negative predictive value of self-reported HPV vaccination status. ResultsThe median age of the 192 males was 19 (IQR 18-20) years. There were 128 males (67%) who had HPV vaccination records documented on either registry. Self-reported HPV vaccination had a sensitivity of 47.7% (95% CI 38.8%-56.7%; 61/128), a specificity of 85.9% (95% CI 75.0%-93.4%; 55/64), a positive predictive value of 87.1% (95% CI 77.0%-93.9%; 61/70), and a negative predictive value of 45.1% (95% CI 36.1%-54.3%; 55/122). ConclusionsSelf-reported HPV vaccination status among Australian gay and bisexual adolescent males underestimates actual vaccination and may be inaccurate for clinical and research purposes

Correlation between HPV16 positivity on anal swab and biopsy-confirmed HSIL caused by HPV16 in the study of the prevention of anal cancer (SPANC)

Background: The majority of anal carcinomas are caused by human papillomavirus (HPV) 16. Identifying which of the high-grade squamous intraepithelial lesions (HSIL) are caused by HPV16 may enable targeted management of those most likely to progress. Methods: In a natural history study of anal HPV-related lesions in gay and bisexual men, HPV typing of all baseline histologic HSIL was performed utilizing laser capture microdissection (LCM). The results were compared with HPV types detected by Cobas 4800 in matched anal swabs. We calculated the performance of Cobas testing in identifying HSIL caused by HPV16. Results: 158 men had both lesion and swab genotyping. Overall 54 men (34%) had HPV16 identified as the causal genotype of at least one HSIL by LCM. 51 of these had HPV16 detected on their anal swabs (sensitivity of 94%). Of the 104 men with non-HPV16 HSIL biopsies, 73 had swabs negative for HPV16 (specificity of 70%). 51 of 82 men with swabs positive for HPV16 had HPV16 positive biopsies (positive predictive value - 62%). 73 of 76 men without HPV16 detected on their swabs had only non-HPV16 HSIL biopsies (negative predictive value (NPV) - 96%). Conclusions: In men with prevalent histologic HSIL, Cobas testing of a concurrent anal swab has high sensitivity for detecting those with HPV16-associated HSIL and a high NPV. HPV testing with the Cobas platform may have a role in the identification and management of those with HSIL caused by HPV16 and who are at highest risk of development of anal carcinoma

Laser capture microdissection-identified HPV genotypes associated with intra-anal high and low-grade intraepithelial lesions

Background: Intra-anal lesions caused by human papillomavirus (HPV) can be classified as low-grade squamous intraepithelial lesions (LSIL) or high-grade squamous intraepithelial lesion (HSIL). The latter can be further subclassified histomorphologically as anal intraepithelial neoplasia (AIN)2 or AIN3. It is not known whether morphological differences within and between these categories are associated with different high-risk HPV (HR-HPV) genotypes. Methods: In a natural history study of anal HPV-related lesions in gay and bisexual men, we identified the presumed causal genotype of all HSIL present at baseline, utilizing laser capture microdissection. For comparison, we also analyzed randomly selected flat LSIL (fLSIL) and negative biopsies. Results: 244 distinct lesions from 175 men were analyzed: 149 AIN3, 64 AIN2 and 31 flat LSIL (fLSIL). HR-HPV genotypes were found in 139 (93.3%) AIN3, 55 (85.9%) AIN2 and 8 (25.8%) fLSIL. HPV16 was found in 59 (39.6%) AIN3, 11 (17.2%) AIN2 and 1 (3.2%) fLSIL. HR-HPV types were not significantly more common in AIN3 than AIN2 (p=0.08) but HPV16 was (p=0.001). HR-HPV genotypes were significantly more common in HSIL than in LSIL (p<0.001) as was HPV16 (p<0.001). None of 24 negative biopsies contained HR-HPV. Conclusions: There were differences observed between the two subdivisions of HSIL, namely AIN3 and AIN2, in proportions of causal HPV16 but not in proportions of HR-HPV overall. As HPV16 is the most carcinogenic HPV subtype, histomorphologic differences within the category of HSIL could have prognostic significance