High prevalence of high-risk HPV genotypes other than 16 and 18 in cervical cancers of Curaçao: implications for choice of prophylactic HPV vaccine

BackgroundCuraçao is a Dutch-Caribbean Island located in a high-risk area for cervical cancer.Prior to introduction of a prophylactic human papillomavirus (HPV) vaccine, knowledge of the prevalence of high-risk HPV vaccine genotypes (HPV16, 18, 31, 33, 45, 52 and 58) in cervical (pre)cancer is required.ObjectiveTo investigate the prevalence of HPV genotypes in invasive cervical cancers (ICC) and cervical intraepithelial neoplasia (CIN) grade 1, 2 and 3 in Curaçao.MethodsParaffin-embedded blocks of 104 cervical cancers (89 squamous, 15 adenocarcinoma), 41 CIN3, 39 CIN2 and 40 CIN1 lesions were analysed for the presence of HPV. Sections were stained by H&E for histopathological evaluation, and DNA was extracted using proteinase K. HPV genotypes were detected using Short PCR Fragment (SPF10) PCR DNA enzyme immunoassay and a Line Probe Assay (LiPA25) .ResultsHPV was found in 92 (88.5%) ICC; 87 (94.6%) had a single HPV infection and 86 (93.5%) were high-risk human papillomavirus (hrHPV)-type positive.The three most common HPV types in ICC were 16 (38.5%), 18 (13.5%) and 45 (6.7%), covering 58.7%.HrHPV vaccine genotypes 16, 18, 31, 35, 45, 52 and 58 were responsible for 73.1% of ICC. For precancerous lesions, the HPV attribution was 85.4% for CIN3, 66.7% for CIN2% and 42.5% for CIN1.ConclusionsOur study, the largest in the Caribbean region in (pre)cancer, shows that the prevalence of HPV-type 16 and 18 in cervical cancer is lower compared with the world population but no differences in prevalence of these two HPV types are seen in precancerous lesions.When considering HPV vaccination in Curaçao, the relatively high contribution of non-HPV 16/18 genotypes in ICC should be taken into account.

Risk, Persistence and Multiplicity of HPV Infections among HIV Negative and HIV Positive Nigerian Women

64 Background: The incidence, prevalence, persistence, and multiplicity of high-risk HPV infection is different between HIV positive and HIV negative women. We examined the association between HIV, prevalent HPV, and persistent HPV infections among women in a prospective cohort in Nigeria. Methods: We enrolled women presenting at cervical cancer screening programs in Abuja, Nigeria, between 2012 and 2014 and collected information on their demographic characteristics, risk factors of HPV infection, and cervical exfoliated cells samples at baseline, 6 month and 12 month follow-up visits. DNA enzyme immunoassay (DEIA) and Roche Linear Array HPV Genotyping Test were used to characterize HPV. Persistent HPV infection was defined as a positive result on 2 consecutive DEIA tests. We used logistic regression models to estimate the association between HIV and risk of HPV infection. Results: Among the 1,020 women enrolled, the mean age (±SD) was 37(8), and 44% and 56% were HIV+ and HIV-, respectively. HPV52 and 35 were the most common HPV types in the study population. The prevalence was 34% for any HPV, 24% for persistent HPV and 9% for multiple HPV infections; these were higher among HIV+ women (p-value <0.001). The multivariate odds ratio (OR) and 95 % CI comparing HIV+ to HIV- women was 6.29 (95% CI 3.64 – 10.87, p-value <0.001) for any high-risk HPV; 6.22 (95% CI 3.02 – 12.83, p-value <0.001) for persistent high-risk HPV; and 6.46 (95% CI 2.69 – 15.52, p-value <0.001) for multiple high-risk HPV infections, Conclusions: HIV infection is associated with increased risk of persistence and multiplicity of low-risk and high-risk HPV infections. These findings may explain, in part, the increased risk of cervical cancer among women with HIV infections. AUTHORS' DISCLOSURES OF POTENTIAL CONFLICTS OF INTEREST: Sally N. Adebamowo No relationship to disclose Toyosi Olawande No relationship to disclose Ayotunde Famooto No relationship to disclose Eileen O. Dareng No relationship to disclose Olayinka Olaniyan No relationship to disclose Richard Offiong No relationship to disclose Clement A. Adebamowo Speakers' Bureau: Merck [Table: see text]

ASSESSMENT OF DIAGNOSTIC TECHNIQUES OF URINARY TUBERCULOSIS

Khalid Ghaleb a,* , Magdy Afifib, Mohamad El-Gohary c aDepartment of Medical Laboratories, Faculty of Applied Medical Science, King Khalid University, Bisha 551, Saudia Arabia bDepartment of Botany and Microbiology, Faculty of Science, Al-Azhar University, Assuit 71524, Egypt cDepartment of Internal Medicine, Faculty of Medicine, Al-Azhar University, Assuit, Egypt The corresponding author e-mail: [email protected] Current Tel: 00966595388496 Saudia, 00201119338055 Egypt The place of the study worked : Department of Botany and Microbiology, Faculty of Science, Al-Azhar University, Assuit 71524, Egypt, e-mail: [email protected] Tel: 00201006554961 Abstract Early diagnosis of active tuberculosis remains an elusive challenge. In addition, one third of the world's population is latently infected with Mycobacterium tuberculosis (Mtb) and up to 10% of infected individuals develop tuberculosis (TB) in their lifetime. In this investigation, the incidence of urinary tuberculosis among renal patients was studied. Three hundreds urine samples were processed for detection of Mtb by Ziehl-Neelson (ZN) smear examination, Lowenstein Jensen (LJ) medium, radiometric BACTEC460 system as well as polymerase chain reaction (PCR) and DNA Enzyme Immunoassay (DEIA) test. Out of 300 urine samples, 2 were positive by both ZN smears and LJ medium with incidence rate of 0.66 %, 3 positive samples by BACTEC460 culture system with incidence of 1%. PCR assay gave more positive results than smear and culture examination (i.e. 8 positive samples with incidence rate of 2.6%). The specificities were 25% for both ZN smears and LJ medium, 37.5% for BACTEC460 culture system, and 100% for PCR test, while sensitivities of all assays were 100%. Thus PCR is a rapid and sensitive method for the early diagnosis of urinary tuberculosis. Keywords: List of abbreviations:Acid Fast Bacilli (AFB)-Base pair (bp)-DNA Enzyme Immunoassay (DEIA) -Extrapulmonary Tuberculosis (EPTB)-International Union Against Tuberculosis (IUAT)-Lowenstein Jensen (LJ)-Mycobacterium tuberculosis (Mtb) -Polymerase Chain Reaction (PCR)-Tuberculosis (TB)-Urinary Tuberculosis (UTB-Urogenital Tuberculosis (UGTB)-Ziehl-Neelson (ZN)

Comparison of Coliforms and Coliphages as Tools for Assessment of Viral Contamination in River Water

ABSTRACT The aim of the study was to evaluate the presence of pathogenic viruses in the Moselle River and to compare the usefulness of thermotolerant coliforms and somatic coliphages as tools for river water quality assessment in terms of viral contamination. Thermotolerant coliforms and somatic coliphages were enumerated by standardized methods in 170 samples of river water drawn from five sampling sites along the Moselle River (eastern France). BGM cell culture and integrated cell culture-reverse transcription-PCR DNA enzyme immunoassay were used to determine the presence of pathogenic viral genome (Enterovirus and Norovirus genogroup II [GGII]) and infectious Enterovirus spp. in 90 1-liter samples. No infectious Enterovirus spp. were isolated, but Enterovirus and Norovirus GGII genomes were detected in 38% of the samples. Norovirus GGII genome was mostly detected in winter, whereas Enterovirus genome was mostly detected in summer and fall. Somatic coliphages appeared to be less sensitive to higher river water temperature than thermotolerant coliforms. Furthermore, the number of river water samples positive for pathogenic viral genome increased with increasing concentration of somatic coliphages, whereas coliform concentration was unrelated to viral genome contamination. Consequently somatic coliphages, which are less sensitive to environmental factors than thermotolerant coliforms in river water, would provide a promising tool for assessment of river water quality in terms of fecal and viral pollution.

Enterotoxigenic Potential ofStaphylococcus intermedius

ABSTRACT Staphylococcal food poisoning (SFP) caused by enterotoxigenic staphylococci is one of the main food-borne diseases. In contrast to Staphylococcus aureus, a systematic screening for the enterotoxins has not yet been performed on the genomic level for the coagulase-positive species S.intermedius. Therefore, the enterotoxigenic potential of 281 different veterinary (canine, n = 247; equine,n = 23; feline, n = 9; other,n = 2) and 11 human isolates of S.intermedius was tested by using a multiplex PCR DNA-enzyme immunoassay system targeting the staphylococcal enterotoxin genes sea, seb, sec,sed, and see. Molecular results were compared by in vitro testing of enterotoxin production by two immunoassays. A total of 33 (11.3%) S.intermedius isolates, including 31 (12.6%) canine isolates, 1 equine isolate, and 1 human isolate, tested positive for the sec gene. In vitro production of the respective enterotoxins was detected in 30 (90.9%) of these isolates by using immunological tests. In contrast, none of 65 veterinary specimen-derived isolates additionally tested and comprising 13 (sub)species of coagulase-negative staphylococci were found to be enterotoxigenic. This study shows on both molecular and immunological levels that a substantial number of S.intermedius isolates harbor the potential for enterotoxin production. Since evidence for noninvasive zoonotic transmission of S. intermedius from animal hosts to humans has been documented, an enterotoxigenic role of this microorganism in SFP via contamination of food products may be assumed.

Development of a Reverse Transcription-PCR–DNA Enzyme Immunoassay for Detection of “Norwalk-Like” Viruses and Hepatitis A Virus in Stool and Shellfish

ABSTRACT Outbreaks of food- and waterborne gastroenteritis are being increasingly reported throughout the world. The analysis of environmental samples by newer diagnostic techniques such as reverse transcription-PCR (RT-PCR) amplification of nucleic acid has begun to identify human enteric viruses (predominantly “Norwalk-like” viruses [NLVs]) as the cause of many of these outbreaks. To streamline NLV detection from environmental samples such as shellfish, we have developed an RT-PCR–oligoprobe amplification and detection method using several new procedures that enable confirmed RT-PCR amplification and product detection in 1 day. The new steps include replacing reverse transcriptase and Taq polymerase with rTth polymerase, a heat-stable enzyme that functions as both a reverse transcriptase and DNA polymerase, in a single-tube, single-buffer, elevated temperature reaction. An internal standard Norwalk virus (NV) RNA control is added to each RT-PCR to identify sample inhibition, and thermolabile uracil N-glycosylase is incorporated into the reaction to prevent PCR product carryover contamination. Finally, RT-PCR-generated amplicons are detected in microtiter wells using virus-specific biotinylated oligoprobes in an enzyme-linked immunosorbent assay-based format. The DNA enzyme immunoassay is based on the capture of PCR product by biotinylated probes fixed onto individual streptavidin-coated wells. Using this method, low levels of NV were detected in stool and both NLV and hepatitis A virus were detected in bivalve mollusks following bioaccumulation. The method also successfully detected NLV in oysters implicated in an outbreak of NLV gastroenteritis. This method dramatically decreases the time needed for analysis and is amenable to automation.