Cervical Swab Samples Research Articles

Antibiotic Susceptibility Pattern of Escherichia coli Isolated from Various Clinical Samples in Duhok City, Kurdistan Region of Iraq

Background: Escherichia coli (E. coli) is one of the most common causative agents of bacterial infections. The emergence of multidrug-resistant E. coli is a major public health threat worldwide. Objectives: This study aimed to determine the antibiotic susceptibility profile of clinical isolates of E. coli from different samples. Methods: A total number of 454 clinical samples, including urine, wound, cervical swab, blood, semen, ascetic, and cerebral spinal fluid samples were collected from patients between January 2017 and February 2020. Then, E. coli was confirmed and susceptibility to different antibiotics was determined using the Vitek-2 compact system. Results: Escherichia coli isolates were more frequent in females (70.7%) than in males (29.3%). In the case of urine samples, E. coli was found to be highly susceptible to ertapenem (97.6%) and imipenem (96.4%) but resistant to ampicillin (87.8%). For wound and cervical swabs, E. coli was 100% resistant to ampicillin and cefepime but 100% sensitive to ertapenem and imipenem. It was found that E. coli isolates from blood samples were 100% resistant to ampicillin, ceftriaxone, and cefoxitin, and around 75% of them were sensitive to ertapenem, ciprofloxacin, and levofloxacin. Finally, E. coli isolated from other clinical samples were highly sensitive to ertapenem, imipenem, levofloxacin, nitrofurantoin, and cefazolin. Conclusions: Escherichia coli isolated from various clinical specimens showed differences in antibiotic sensitivity patterns, with high resistance to commonly used antibiotics. The most effective antibiotics against E. coli isolates were ertapenem, imipenem, and nitrofurantoin. However, the clinical isolates of E. coli displayed high resistance rates to ampicillin, ceftriaxone, and cefepime. Therefore, it is proposed to perform antibiotic sensitivity testing by physicians to select the most effective antibiotics.

Evaluation of microRNA-20, -21 and -143 expression in human papilloma virus induced premalignant and malignant cervical lesions

Cervical cancer (CC) is the fourth most prevalent cancer worldwide. The deprivation of screenin gprogram implementation and inefficient conventional treatments of CC had led to diagnosis of disease in advanced stages and poor prognosis due to gradual increase in resistance to chemotherapy. Therefore, identification of CC molecular targets had become a primary task in order to improve detection, prognosis and development of cervical cancer targeted therapy. MicroRNAs (miRNA) have been validated as key players in cell physiology and alterations in miRNA expression were associated with cancer progression and therapy response. This study was designed to determine the expression of three microRNA molecules involved in human papilloma virus (HPV) induced cervical disease progression in Iraqi women for early detection of preneoplastic and neoplastic cervical lesions. A total of fifty cervical swab samples were collected from women who presented with cervical abnormalities and thirty samples from volunteers healthy women as a control group. Levels of miRNA expression were measured by two steps reverse transcription polymerase chain reaction (RT-PCR) technique. Results showed a significant increase in the expression levels of miRNA-21 and miRNA-20 (P = 0.0001 and 0.048, respectively) in HPV positive patients in comparison with HPV negative and healthy controls. Whereas, miRNA-143 showed a dichotomous expression pattern as being transiently upregulated (P = 0.0001) in preneoplastic lesions (CIN-II and CIN-III) and down regulated at cervical carcinoma. These data suggest that overexpression of cellular miRNA-21, -20 and -143 may be related to infection with high risk HPV and could be a biomarker and a useful therapeutic target.

Chlamydia trachomatis and Human Papillomavirus Infection in Women From Southern Hunan Province in China: A Large Observational Study.

Chlamydia trachomatis and human papillomavirus (HPV) are the most common pathogens of sexually transmitted infections (STIs), which can increase the risk of cervical cancer and infertility. The purpose of this study was to evaluate the prevalence, genotype and risk factors of C. trachomatis and/or HPV infection in women attending the annual physical examination, assistant reproductive treatment and visiting the gynecology clinics from Southern Hunan province in China. Cervical-swab samples were collected from 5006 participants. We found that the overall prevalence of C. trachomatis, HPV infection and C. trachomatis/HPV coinfection was 4.7% (236/5006), 15.5% (778/5006) and 1.2% (59/5006), while the prevalence of asymptomatic infection of that was 3.8% (38/1006), 10.8% (109/1006) and 0.6% (6/1006), respectively. Furthermore, 25.0% (59/236) of C. trachomatis infection and 7.6% (59/778) of HPV infection were attributable to C. trachomatis and HPV coinfection. C. trachomatis and HPV infection were more often observed in young women of less than 25 years (10.4% and 21.3%, respectively) and in the outpatients from gynecology clinics (5.2% and 18.0%, respectively). Of note, a higher prevalence of C. trachomatis infection was observed in HPV-positive women (7.6%) than HPV- negative ones (4.2%), and vice versa. The top three C. trachomatis genotypes were E (1.4%), F (1.1%) and J (0.8%), and the counterparts of HPV genotypes were HPV52 (4.2%), HPV16 (2.3%) and HPV58 (2.2%), respectively. Among the 151 outpatients with colposcopy data, HPV infection was associated with severe cervical lesions with OR of 15.86 (95% CI 3.14–80.0, P < 0.001) while C. trachomatis infection was more likely associated with a low grade colposcopy impression (OR = 3.25, 95% CI: 1.22–8.65, P = 0.018). Our data highlight the high prevalence of asymptomatic C. trachomatis and HPV infection, particularly among women of <25 years. The two pathogens may serve as mutual risk factors to increase the risk of infections and cervical lesions. Widespread implementation of HPV and C. trachomatis screening programs, especially for young women, would be an effective strategy to relieve the burden of sexually transmitted infections.

Genotyping of Human Papillomavirus in Cervical Squamous Intraepithelial Lesions in Mexican Women.

Approximately 40 genotypes of the human papillomavirus (HPV) have been identified in cervical mucosa. In particular, HPV-16 and HPV-18 have been associated with cervical neoplasia. Squamous intraepithelial lesions (SILs) are precursors of cervical cancer. This study aimed to identify the HPV by genotype in SILs using a linear array genotyping test in a population in Mexico. We performed a cross-sectional study of 129 female patients with or without SILs, as determined by colposcopy, who completed a risk factor questionnaire. Cervical swab samples were obtained and genotyped using a Linear Array HPV Genotyping assay. Forty-nine (37.98%) samples were positive for HPV, and 24 genotypes were identified among these samples. The most common genotype was HPV-16. Twelve genotypes were found in both high- and low-grade SILs (HPV-6, 16, 31, 39, 51, 52, 53, 58, 59, 61, 67, and 84), of which seven were high-risk SILs (HPV-16, 31, 39, 51, 52, 58, and 59). Among the populations studied, the most frequent genotype was HPV-16, multiple infections were found, and four patients without injury tested positive for HPV.

Methylation of CpG 5962 in L1 of the human papillomavirus 16 genome as a potential predictive marker for viral persistence: A prospective large cohort study using cervical swab samples.

Several studies have demonstrated that the viral genome can be methylated by the host cell during progression from persistent infection to cervical cancer. The aim of this study was to investigate whether methylation at a specific site could predict the development of viral persistence and whether viral load shows a correlation with specific methylation patterns. HPV16‐positive samples from women aged 20–29 years (n = 99) with a follow‐up time of 13 years, were included from a Danish cohort comprising 11 088 women. Viral load was measured by real‐time PCR and methylation status was determined for 39 CpG sites in the upstream regulatory region (URR), E6/E7, and L1 region of HPV16 by next‐generation sequencing. Participants were divided into two groups according to whether they were persistently (≥ 24 months) or transiently HPV16 infected. The general methylation status was significantly different between women with a persistent and women with a transient infection outcome (P = .025). One site located in L1 (nt. 5962) was statistically significantly (P = .00048) different in the methylation status after correction using the Holm‐Sidak method (alpha = 0.05). Correlation analyses of samples from HPV16 persistently infected women suggest that methylation is higher although viral load is lower. This study indicates that methylation at position 5962 of the HPV16 genome within the L1 gene might be a predictive marker for the development of a persistent HPV16 infection.

Changes in DNA Level of Oncogenic Human Papillomaviruses Other Than Types 16 and 18 in Relation to Risk of Cervical Intraepithelial Neoplasia Grades 2 and 3.

Epidemiologic data addressing clinical relevance of viral load fluctuation of oncogenic types other than human papillomavirus (HPV) types 16 and 18 are limited. A type-stratified set of infections by non-HPV16/18 oncogenic types that were detected at ≥2 visits was randomly selected from women who were enrolled in a clinical trial and followed every 6 months for 2 years for detection of HPV and cervical intraepithelial neoplasia grades 2 and 3 (CIN2/3). Type-specific viral load was measured on both first and last HPV-positive cervical swab samples. CIN2/3 was initially confirmed at the last HPV-positive visit for 67 of 439 infections. The increase in risk of CIN2/3 was associated with high, relative to low, viral load at both first and last positive visits [ORadjusted = 3.67; 95% confidence interval (CI), 1.19-11.32] and marginally associated with a change of viral load from low to high levels (ORadjusted = 3.15; 95% CI, 0.96-10.35) for infection by species group alpha-9 non-HPV16 oncogenic types but not species group alpha-5-7 non-HPV18 oncogenic types. Among women with an initial diagnosis of CIN2/3 at the first positive visit, CIN2/3 was more frequently redetected at the last positive visit for infections with, compared with without, high DNA load of species group alpha-9 non-HPV16 oncogenic types at both visits (P exact = 0.04). In agreement with data on baseline viral load, the viral load change-associated risk of CIN2/3 differs by HPV species groups. These findings underscore the importance of distinguishing species groups in future studies of clinical relevance of HPV DNA load.

Composite Analysis of the Virome and Bacteriome of HIV/HPV Co-Infected Women Reveals Proxies for Immunodeficiency.

The human cervical microbiome is complex, and its role in health and disease has just begun to be elucidated. In this study, 57 cervical swab samples from 19 HIV/HPV co-infected women were analyzed for both virome and bacteriome composition. Virome analysis focused on circular DNA viruses through rolling circle amplification followed by next-generation sequencing (NGS). Data were assigned to virus families and genera, and HPV types were identified. NGS data of bacterial 16S from a subset of 24 samples were assigned to operational taxonomic units and classified according to vaginal microbiome community state types (CSTs). Four viral families were found: Papillomaviridae, Anelloviridae, Genomoviridae, and Herpesviridae. Papillomavirus reads were more abundant in women with premalignant cervical lesions, which were also strongly associated with multiple (≥3) high-risk HPV infection. Anellovirus read abundance was negatively correlated with host CD4+ T-cell counts. The bacteriome revealed the presence of CST III and CST IV, and women with ≥1% frequency of genomovirus or herpesvirus reads displayed an increased risk of carrying CST IV. By characterizing the composition of the cervical circular DNA viruses and the bacteriome of HIV/HPV co-infected women, we identified putative interactions between these two microorganism communities and their associations with patients’ clinical characteristics, notably immunodeficiency status.

Comparison of urine, self-collected vaginal swab, and cervical swab samples for detecting human papillomavirus (HPV) with Roche Cobas HPV, Anyplex II HPV, and RealTime HR-S HPV assay

BackgroundHuman papillomavirus (HPV) is well established as the main cause of cervical cancer. Non-invasive self-collected urine and vaginal sampling have the potential advantage of increasing patient compliance with cervical cancer screening. MethodsSelf-collected vaginal and urine samples and clinician-collected cervical samples were collected from 101 patients, including 84 patients with high grade squamous intraepithelial lesion and 17 patients with benign ovarian disease. Each sample was evaluated with RealTime HR-S HPV, Anyplex™ II HPV, and Cobas® HPV assays. The concordance of urine and of self-collected vaginal samples with cervical samples was assessed using the kappa (k) statistic. ResultsIn any high-risk HPV (hrHPV), the concordance of self-collected vaginal and urine samples compared to cervical samples was moderate (k 0.49–0.58) and fair to moderate (k 0.33–0.51), respectively. In HPV 16/18, the concordance of vaginal and urine samples compared to cervical samples was almost perfect (k 0.81–0.86) and moderate to substantial (k 0.59–0.63), respectively. Among the three methods for HPV detection, RealTime HR-S showed the highest concordance with vaginal (k: any hrHPV 0.58, HPV 16/18 0.86) and urine samples (k: any hrHPV 0.51, HPV 16/18 0.63) compared to cervical samples. ConclusionHPV tests using self-collected vaginal samples and urine showed substantial and moderate agreement compared with cervical samples, respectively, although HPV tests using these samples were still inferior to clinician-collected cervical samples. Further research is needed on the clinical performance of HPV testing using urine and self-collected vaginal samples as the screening method.

Prevalence of Chlamydia trachomatis Infection Diagnosed by Polymerase Chain Reaction in Female Sex Workers in a Northern Mexican City.

PurposeWe aimed to determine the association between Chlamydia trachomatis infection and female sex work, and the association between sociodemographic, obstetric, and behavioral characteristics of female sex workers and C. trachomatis infection.MethodsThrough a case–control study design, we studied 201 female sex workers and 201 age-matched women without sex work in Durango City, Mexico. C. trachomatis DNA was detected in cervical swab samples using polymerase chain reaction.ResultsC. trachomatis DNA was detected in 32 (15.9%) of the 201 cases and in 6 (3.0%) of the 201 controls (odds ratio [OR] = 6.15; 95% confidence interval [CI]: 2.5–15.0; P < 0.001). The frequency of infection with C. trachomatis in female sex workers did not vary (P > 0.05) regardless of the history of pregnancies, deliveries, cesarean sections, or miscarriages. Regression analysis of the behavioral characteristics showed that infection with C. trachomatis was associated only with consumption of alcohol (OR = 2.39; 95% CI: 1.0–5.71; P = 0.04). Conclusions: We conclude that C. trachomatis infection is associated with female sex work in Durango City, Mexico. This is the first age-matched case–control study on the prevalence of C. trachomatis infection in female sex workers in Mexico using detection of C. trachomatis DNA in cervical samples.

Investigation of human papillomavirus prevalence in married women and molecular characterization and phylogenetic analysis of the virus.

ObjectiveHuman papillomaviruses (HPVs) are among the agents responsible for infection and cancer of the skin and mucous membranes in the human body. The aim of this study was to investigate the frequency and type distribution of HPVs in married female patients with gynecological complaints, who had visited the Maternity Hospital in Erzurum, Turkey.MethodsIn this study, 263 cervical swab samples were taken from married women using the Pap smear method and were investigated for positive reactivity against HPV. The L1 gene region of HPV was investigated using molecular methods. For this purpose, polymerase chain reaction (PCR) assays and sequence analysis of positive samples were performed. Phylogenetic analyses were performed using a bioinformatics approach after sequencing.ResultsHPV-DNA was detected in 17 (6.5%) samples. Highest positive reactivity to HPV-DNA was found in the 35–44 age group at 9.2%. Fourteen out of seventeen positive samples were included in the phylogenetic analysis. All isolates clustered in the Alphapapillomavirus genus. Six samples were found to be HPV 70 positive, four were HPV 16 positive, and the rest were HPV 54, 72, 81, and 114 positive. When genotyping data were evaluated according to the risk group, we found that 28.6% of the 14 samples were found to be high risk-HPV, and 71.4% were low risk-HPV.ConclusionsAs per our knowledge, this is the first report on the phylogenetic analysis of HPV genotypes isolated from women in Turkey. The prevalence of low- and-high risk HPV was determined in married women in Erzurum, and these results contribute to the epidemiological data on the distribution of HPV types for this region.

The Molecular Detection of HPV Infection in samples of Iraqi Women with Abnormal cervical Smears

Human papillomavirus (HPV) types 16 and 18 cause almost 70% of cervical cancer cases worldwide. Recently, testing for high-risk HPV types have been adopted by clinical practices for the early detection of cervical cancer in conjunction with cytology tests. Cervical swab samples were collected at the Outpatient Gynecology department of Baghdad Teaching Hospital. These samples consisted of a patient group of 50 samples, and a healthy control group of 10 samples. A papanicolaou test (abbreviated as a Pap test) was also performed for each woman to examine the epithelial cells of both the endocervix and the upper vaginal region. Total DNA (genomic, mitochondrial, and viral) was extracted from cervical swab samples for molecular studies. HPV DNA testing was first done by using Real-Time PCR technology to target the L1 region of HR-HPV with specifically designed primers. This was followed by using AmpliSens kit for specific detection and genotyping of HPV16 and 18 with multiplex Real-Time PCR. The results of RT-PCR detection revealed that out of 16 samples detected with high-risk HPV, 5 samples were shown to be infected with HPV-16 and 5 samples were shown to be infected with HPV-18. These results show a significant relationship between the histological outcome of the patient and persistent HPV infection.

Impact of bacterial traces belonging to the Enterobacteriaceae on the prevalence of Chlamydia trachomatis in women visiting a community hospital in Japan.

We explored the bacteria present in the vaginal microbiota facilitating the prevalence of Chlamydia trachomatis in women visiting a community hospital in Sapporo, Japan, by amplicon sequencing. A total of 273 cervical swab samples were collected, and bacterial vaginosis was evaluated in all specimens by assessment of the Nugent score. In 16 of the samples, bacterial 16S rDNA could not be detected and they were therefore omitted from subsequent experiments (n=257). A significant negative correlation was observed between the Nugent scores and the amount of Lactobacillus 16S rDNA. Among the 257 samples, chlamydial plasmid was detected in 20 samples and was used for amplicon sequencing. No significant association between the Nugent score and the prevalence of C. trachomatis was detected. Based on the results of chlamydial plasmid detection and the Nugent score, chlamydia-negative samples (n=27) were randomly selected. Finally, the number of operational taxonomic units (OTUs) obtained from amplicon sequencing was compared between chlamydia-positive (n=20) and -negative samples (n=27), revealing that a significant difference was only detected for the OTU numbers of Enterobacteriaceae between the C. trachomatis-positive and -negative groups. However, almost all of the samples utilized for amplicon sequencing failed to grow on MacConkey agar plates and produce indole. Taken together, we concluded that traces of bacteria, not live bacteria, belonging to the Enterobacteriaceae indicated the flow of bacteria through the anogenital route along with gut indole, and the resulting impact on the prevalence of C. trachomatis in the cervicogenital tract of women in Japan.

6 - Pilot study of markers for high-grade anal dysplasia in a southern cohort from the Women's Interagency HIV Study (WIHS)

Background Anal cancer rates have increased in HIV+ women. Factors associated with anal cancer precursor high-grade squamous intraepithelial lesions (HSIL) in HIV+ and at-risk-HIV- women were assessed. Methods HIV+ and HIV- women from the Atlanta WIHS cohort were enrolled in a cross-sectional pilot study. All anal (AS) and cervical (CS) swab samples were analyzed for HPV-genotyping (Linear-Array® HPV-Genotyping Test, LA-HPVGT) and FAM19A4 and microRNA-124-2 promoter methylation. All women underwent high resolution anoscopy with biopsy of suspicious lesions and anal cytology (AC) collection. Logistic regression was conducted with anal HSIL histology (A-HSIL) as the dependent variable. Results 75 women were enrolled: 52(69%) were HIV+ with 3/4 having undetectable viral load, 64(86%) Black, with mean age 49. 44(59%) AC samples were ASCUS/+hrHPV or higher, 38(51%) of all AS were +hrHPV by LA-HPVGT.13 anal biopsies were confirmed A-HSIL. 69(95%) AS and 19(26%) CS tested positive for hypermethylation, respectively. A-HSIL model included ASCUS/+hrHPV or greater on AC and cervical hypermethylation as covariates (Table 1). AS hypermethylation was not associated with A-HSIL. Conclusions Our results suggest AC with hrHPV testing and/or cervical gene promoter hypermethylation measurements as promising non-invasive screening strategies for A-HSIL in both HIV+ and HIV- women. Lack of association between AS hypermethylation and A-HSIL may reflect characteristics of the anal milieu and warrants further investigation.

Detection and genotyping of human papillomavirus (HPV) in HIV-infected women and its relationship with HPV/HIV co-infection.

HPV have been identified as high-risk and low-risk, depending on their association with the development of cancer. HPV infections can be facilitated by co-infection with HIV. Here, we investigated HPV prevalence and genotypes and the risk factors affecting HPV/HIV co-infection. Forty HIV-positive patients had 80 cervical swab samples collected in 2 consecutive years. Polymerase chain reaction and DNA direct sequencing were used to perform HPV genotyping. Statistical analyses were performed regarding risk factors for HPV/HIV co-infection and the occurrence of cervical lesions. HPV DNA was detected in 59 samples (73.75%), and high-risk HPVs were predominant (59.3%). The most prevalent type was HPV56 (17%), followed by HPV16 (15.3%). Patient age did not affect the risk of cervical cancer (P = .84) or HPV prevalence in different years (P = .25/P = .63). CD4 count also did not affect the risk for cervical lesions in the tested samples (P = .15/P = .28). Although the HIV viral load was not correlated with an increase in cervical lesion detection in the first group of analyzed samples (P = .12), it did affect cervical cancer risk in the group of samples analyzed in the following year (P = .045). HIV-infected patients presented a high prevalence of HPV co-infection, and HPV16 and HPV56 were the most prevalent genotypes. Considering this, it is possible that immunodeficiency can contribute to increased susceptibility to HPV56 infection in HIV-infected patients. The association between HIV viral load and the lesions also confirmed the importance of monitoring HIV/HPV co-infected patients with high HIV viral loads.

Accuracy of syndromic management in targeting vaginal and cervical infections among symptomatic women of reproductive age attending primary care clinics in Dakar, Senegal.

To assess the effectiveness of the WHO syndromic algorithm in the management of vaginal discharge among women of reproductive age in Dakar. Cross-sectional study of consecutive female patients (aged 18-49 years) presenting with vaginal symptoms at six selected study sites in Dakar; of these, 276 patients were included in the analysis. Vaginal and cervical swab samples were collected and analysed to establish an aetiological diagnosis of any infection. Syndrome-based diagnosis was compared with the laboratory results to evaluate its accuracy based on sensitivity, specificity and positive and negative predictive values. The degree of agreement between the two approaches was assessed using the Cohen's kappa concordance analysis. Overall prevalence of vaginal infections was 56.9% (157/276); 5.4% (15/276) of the patients had cervical infection. Using the syndromic approach, 51% of patients were correctly managed for Trichomonas vaginalis (TV)/Gardnerella vaginalis (GV); 61% for Candida albicans (CA) and 54% for Chlamydia trachomatis (CT)/Neisseria gonorrhoea (NG) infections. Consequently, 31% of patients with TV/GV, 51% with CA and 53% with CT/NG infections would have missed treatment. Further, the kappa value was <0.20, indicating that there was no agreement or only slight agreement between the syndromic approach and laboratory-based diagnosis. This study highlights the limitations of the applicability of the WHO syndromic approach in settings with low prevalence of sexually transmitted infections (STIs) and calls for affordable and accurate rapid tests for STIs.

Cervical biomarkers as predictors of successful induction of labour by Foley catheter

The prediction of successful labour induction is difficult, indicating a need for a biomarker test. Little is known about the effect of Foley catheter (FC) induction on biochemical mediators in the cervix, such as the insulin-like growth factor binding protein-1 (IGFBP-1), matrix metalloproteinases (MMP) and their inhibitors (TIMP). We enrolled 35 nulliparous women with singleton pregnancies, intact amniotic membranes and cephalic presentation ≥40 gestational weeks scheduled for labour induction by FC. Serial cervical swab samples were collected at FC insertion and expulsion. The concentrations of IGFBP-1, PhIGFBP-1, MMP-8, MMP-2, MMP-9, TIMP-1 and TIMP-2 were analysed. The IGFBP-1 and phIGFBP-1 concentrations increased during the FC-induced cervical ripening. In contrast, MMP-8 and MMP-9 concentrations decreased. However, these changes did not predict the outcome of the labour induction, thus appearing not suitable for clinical use.Impact statementWhat is already known on this subject? During cervical ripening, various constituents interact in a complex network. Insulin-like growth factor binding protein-1 (IGFBP-1), matrix metalloproteinases (MMP) and their tissue inhibitors (TIMP) appear to play a role in cervical ripening. The mechanism of Foley catheter on cervical ripening consists of direct mechanical stretching of the cervix and lower uterine segment, and the stimulation of local secretion of endogenous prostaglandins.What do the results of this study add? This study investigated the role of cervical biochemical mediators during Foley catheter-induced cervical ripening, and their predictive value in a successful labour induction and vaginal delivery. The IGFBP-1 and phosphorylated IGFBP-1 concentrations increased, whereas MMP-8 and MMP-9 concentrations decreased during the Foley catheter-induced cervical ripening in nulliparous women. However, these changes did not predict the outcome of labour induction, thus appearing not suitable for clinical use.What are the implications of these findings for clinical practice and/or further research? Prediction of a successful labour induction is difficult, indicating a need for a biomarker test. Future studies with larger data are needed for investigating the role of these cervical biomarkers in successful labour induction, and in developing a future bedside a screening tool for clinical use.

A Multiplexed Human Papillomavirus 16 and 18 Diagnostic Chip for Cervical Cancer in Limited-Resource Settings

Abstract 16 Purpose Despite being treatable, cervical cancer is responsible for 300,000 deaths annually and is the second most frequent cause of cancer-related death worldwide. The burden of cervical cancer disproportionately falls on developing countries, where 87% of deaths occur from limited or no resources for screening. Of cervical cancer cases, 99% are caused by human papillomavirus (HPV), with more than 70% of cases from HPV genotypes 16 and 18. We developed a low-cost cervical cancer diagnostic chip to detect HPV 16 and 18 DNA quickly and reliably in limited-resource settings (LRSs). Methods Unlike conventional HPV diagnostics that use polymerase chain reaction and resource-intensive equipment, our chip used isothermal—one temperature—amplification, specifically loop-mediated amplification, and required only a heat source, which made it suitable for LRSs. Amplified DNA was visually detected on a lateral flow strip, which is similar to a pregnancy test, providing a simple yes or no readout. Specific line patterns indicated the presence of HPV 16, 18, both, or neither. Each chip was single use and self-contained to be easily disposed of as biohazard waste, which reduced the risk of contamination and false positives. Results The loop-mediated amplification assay was optimized to detect HPV 16 and 18 simultaneously. Sensitive and specific amplification of cloned HPV 16 and 18 DNA was achieved and confirmed via specific restriction enzyme digests. The lower limit of detection was 104 and 103 copies for HPV 16 and 18, respectively. Clinical samples, namely discarded cervical swab samples, were also tested with the chip. Results were comparable with the gold standard of polymerase chain reaction, which proves that the chip is feasible for clinical samples. Time to result was less than 1 hour, making the chip appropriate for LRSs. Conclusion Our multiplexed HPV 16 and 18 diagnostic chip is clinically relevant and provides a much-needed screening method for LRSs. The chip will increase access to screening for a treatable cancer and provide a faster route to treatment as well as a decrease in deaths from cervical cancer. AUTHORS' DISCLOSURES OF POTENTIAL CONFLICTS OF INTEREST No COIs from the authors.

Comparison of the Tellgenplex HPV DNA test with the PCR-reverse dot blot assay for human papillomavirus genotyping

Objective: To access the performance of the Tellgenplex human papillomavirus (HPV) DNA test compared to the polymerase chain reaction-reverse dot blot (PCR-RDB) assay for the HPV genotyping. Methods: Sixty cervical swab samples were genotyped by the Tellgenplex HPV DNA test and the PCR-RDB assay. The Tellgenplex HPV DNA test and the PCR-RDB assay can detect 26 and 23 HPV genotypes, respectively. Each sample showed discrepancy was genotyped using sequencing. Results: The percent agreement between the two tests ranged from 83.3% to 100.0% according to different genotype. This showed perfect agreement (>0.81) for high-risk HPV genotypes (35, 39, 45, 53, 56, 59, 66, 68, and 82), substantial agreement (>0.65) for high-risk HPV genotypes (16, 18, 33, 52, and 58) and low-risk HPV genotype 43 between the two assays by the kappa analysis. The positive rates of the two assays for frequent HPV genotypes (16, 35, 39, 45, 52, 53, 58, 59, 66, and 82) were not statistically different, but the PCR-RDB assay showed higher positive rates than the Tellgenplex HPV DNA test for HPV genotypes 81 (P

Chlamydia trachomatis urogenital infection and associated risk factors among university students in Croatia

O bjective − The study aimed to determine the prevalence of Chlamydia trachomatis infection among university students in Croatia and to determine risk factors associated with chlamydia infection. M aterial and methods − The study was conducted as an opportunistic screening of asymptomatic sexually active students attending routine health examinations during their first year of studying. The first void urine samples from males, cervical swab or first void urine samples from females and data regarding life style, sexual and other health-risk behaviour were collected and analysed. R esults − Overall prevalence of chlamydia infection was 4.9%, with the testing response rate of 29.5%. The only identified risk factor was female sex. Female students preferred urine sample over cervical swab sample and avoided gynaecologist examination. Conclusion − Opportunistic screening is a feasible strategy to diagnose asymptomatic chlamydia infection among university students in Croatia. Results may contribute to decision making on the form of chlamydia control policy, including choice of screening settings and specimens suitable for target population.

Comparison of human papillomavirus (HPV) detection in urine and cervical swab samples using the HPV GenoArray Diagnostic assay.

Human papillomavirus (HPV) is the leading cause of cervical cancer. Urine-based HPV testing offers a simple and non-invasive method because of its increasing acceptance. A total of 164 pairs of cervical swab and urine samples from Thai women who underwent cervical cancer screening were used for HPV testing with HPV GenoArray Diagnostic Kits. The overall concordance percentage for HPV detection in the cervical swab and urine samples was 65.2%. The HPV genotypes most commonly detected were HPV16 and HPV18. An analysis of the urine samples and a second analysis of the cervical swab samples showed that the differences in the overall HPV detection rate between women with normal and abnormal cytology were not significant (p > 0.05). Urine samples processed with the GenoArray assay is an alternative for women who decline to undergo Pap smear even though it is not ideal as the first-line screening option.