scholarly journals Bacterial etiology of sexually transmitted infections at a STI clinic in Ghana; use of multiplex real time PCR

2016 ◽  
Vol 50 (3) ◽  
pp. 142-148
Author(s):  
Augustina A Sylverken ◽  
Ellis Owusu-Dabo ◽  
Denis D Yar ◽  
Samson P Salifu ◽  
Nana Yaa Awua-Boateng ◽  
...  
Keyword(s):  
Sexually Transmitted Infections ◽  
Real Time ◽  
Real Time Pcr ◽  
Pathogen Detection ◽  
Clinical Manifestations ◽  
Diagnostic Tools ◽  
Transmitted Infection ◽  
Sexually Transmitted ◽  
Asymptomatic Patients ◽  
Syndromic Approach

Background: Most sexually transmitted infection (STI) management efforts focus on the syndromic approach to diagnose and treat patients. However, most women with STIs have been shown to be entirely asymptomatic, or if symptoms exist, are often missed when either clinical or conventional bacteriologic diagnostic tools are employed.Methods: We assessed the performance of a multiplex real time PCR assay to describe other potential pathogens that could be missed by conventional bacteriological techniques in 200 women attending a routine STI clinic in Kumasi, Ghana.Results: Although a total 78.00% of the women were asymptomatic, 77.1% of them tested positive for at least one bacterial STI pathogen. Mycoplasma genitalium was the most commonly detectable pathogen present in 67.5% of all women. Of those testing positive, 25.0% had single infections, while 38.0% and 19.5% had double and triple infections respectively. Altogether, 86.54% and 90.91% of the symptomatic and asymptomatic women respectively tested positive for at least one pathogen (p<0.05). There were no significant associations (p<0.05) between the clinical manifestations of the symptomatic women and the pathogens detected in their samples.Conclusions: Our study confirmed the importance of complementing the syndromic approach to STI management with pathogen detection and most importantly recognise that STIs in women are asymptomatic and regular empirical testing even for both symptomatic and asymptomatic patients is critical for complete clinical treatment.Funding: EOD (Ellis Owusu-Dabo Research working group, KCCR)Keywords: Etiology, Syndromic, Sexually Transmitted Infections, Multiplex real time PCR

scholarly journals Direct detection of microorganisms responsible for the sexually transmitted infection on the mobile real-time PCR device without treating in advance

2021 ◽  
Author(s):  
Masaaki Muraoka ◽  
Kazunori Sohma ◽  
Osamu Kawaguchi ◽  
Mikio Mizukoshi
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Limit Of Detection ◽  
Direct Detection ◽  
Treponema Pallidum ◽  
Nucleic Acid Amplification ◽  
Direct Pcr ◽  
Transmitted Infection ◽  
Sexually Transmitted ◽  
Ct Value

ABSTRACTAs WHO reported, four curable STIs-chlamydia, gonorrhoea, syphilis and trichomoniasis occur more than 1 million per each day globally almond 2016. For this reason, it is important to control these STIs, one of which is “to detect”. The general methods in order to detect STIs are nucleic acid amplification tests (NAATs). One of the reasons why NAATs are utilized in many tests is that it is possibly to be more sensitive than other test. However, there needs to treat extraction of nucleic acids in advance and amplify specific regions by NAATs, and hence it must take much labour and much time. In this work, for Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and Treponema pallidum (TP) which is each etiological agent of chlamydia, gonorrhoea and syphilis, we evaluate and propose “quicker and simpler” NAATs. Specifically, utilizing mobile real-time PCR device “PCR1100” and PCR reagent kit “KAPA3G Plant PCR Kit”, it was considered whether real-time direct PCR could be performed or not without treating DNA extraction in advance so-called “direct”.As a result, firstly, we established that real-time direct PCR could be performed in all of CT, NG, and TP, and moreover, each Ct value correlated with the concentration of each organism similarly to detection of genome DNA (each correlation coefficient R2 > 0.95). Moreover, each assay demonstrated a limit of detection (LOD) of the follows; CT was 10^0.86 = 7.24 IFU/reaction, NG was 10^-0.19 = 0.65 CFU/reaction, and TP was 10^1.4 = 25.1 organisms/reaction. However, it appeared the sensitivity was a little low, especially for CT and TP.Secondly, we found that even as without treating sample in advance, the time of detection was required more less 15 minutes at any of case, which was very quick compared with other current methods for real-time PCR. Additionally, compared with other commercial devices, it was easier to operate the PCR1100 device, for example, start, analysis of Ct value.In conclusion, the present study has demonstrated that it is possible for real-time direct PCR to perform with combination of the PCR1100 device and the PCR reagent kit in 3 kinds of microorganisms-CT, NG and TP. Furthermore, we propose “quicker and simpler” methods for NAATs, which it would not take labour and time. Further studies are needed in order to contribute to control STIs.

scholarly journals Sexually Transmitted Infections Detected by Multiplex Real Time PCR in Asymptomatic Women and Association with Cervical Intraepithelial Neoplasia

2018 ◽  
Vol 40 (09) ◽  
pp. 540-546 ◽  
Author(s):  
Luiza Lima ◽  
Carolina Hoelzle ◽  
Renata Simões ◽  
Maria Lima ◽  
Jordana Fradico ◽  
...  
Keyword(s):  
Multivariate Analysis ◽  
Cervical Intraepithelial Neoplasia ◽  
Sexually Transmitted Infections ◽  
Real Time ◽  
Ureaplasma Urealyticum ◽  
Significant Risk ◽  
Intraepithelial Neoplasia ◽  
Sexually Transmitted ◽  
Asymptomatic Patients ◽  
Asymptomatic Women

Objective To determine the frequency of sexually transmitted infections (STIs) in asymptomatic women and the association of STIs with cervical intraepithelial neoplasia (CIN). Methods A cross-sectional study was performed, enrolling women examined in a general gynecology clinic and in a colposcopy referral center from October 2014 to October 2015. The colposcopy group consisted of 71 women, and the general gynecology group consisted of 55 women. Cervical samples were collected for cervical cytology and a multiplex real-time polymerase chain reaction (PCR) was developed to detect human papillomavirus (HPV) and the STIs caused by the following microorganisms: Chlamydia trachomatis, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum, and Neisseria gonorrhoeae. A multivariate analysis was performed by logistic regression, considering the significance level of 0.05. Results The general frequency of STIs was: 46.8% (HPV); 27.8% (C. trachomatis); 28.6% (M. genitalium); 0.8% (M. hominis); 4.8% (U. urealyticum); and 4.8% (N. gonorrhoeae). The significant risk factors for CIN were: HPV infection (odds ratio [OR] = 2.53; p = 0.024); C. trachomatis (OR = 3.04; p = 0.009); M. genitalium (OR = 2.37; p = 0.04); and HPV and C. trachomatis coinfection (OR = 3.11; p = 0.023). After the multivariate analysis, a significant association was found between HPV and CIN (OR = 2.48; 95% confidence interval [95%CI]: 1.04–5.92; p = 0.04); and between C. trachomatis and CIN (OR = 2.69; 95%CI: 1.11–6.53; p = 0.028). Conclusion The frequency of STIs was high in asymptomatic patients. Infections by HPV and C. trachomatis were independently associated with the presence of CIN. The high frequency of STIs in asymptomatic women suggests the need for routine screening of these infections.

scholarly journals A comparison of oligonucleotide-based microarray and real-time PCR for the detection of sexually transmitted infections

2013 ◽  
Vol 7 (1) ◽  
pp. 68-74 ◽  
Author(s):  
Gyeong-In Lee ◽  
Jong Pil Yoen ◽  
Jin Seok Kang ◽  
Seung Yong Hwang ◽  
Yu-Min Hong ◽  
...  
Keyword(s):  
Sexually Transmitted Infections ◽  
Real Time ◽  
Real Time Pcr ◽  
Sexually Transmitted

Evaluation of VIASURE Sexually Transmitted Diseases Real Time PCR Detection kit (CerTest Biotec) for the diagnostic of sexually transmitted infections

2021 ◽  
Vol 39 (5) ◽  
pp. 229-233
Author(s):  
Cristina Casañ López ◽  
Belén Rivaya Sánchez ◽  
Gema Fernández Rivas ◽  
Águeda Hernández Rodríguez ◽  
Adrián Antuori Torres ◽  
...  
Keyword(s):  
Sexually Transmitted Infections ◽  
Sexually Transmitted Diseases ◽  
Real Time ◽  
Real Time Pcr ◽  
Pcr Detection ◽  
Sexually Transmitted

scholarly journals Single gene targeted nanopore sequencing enables simultaneous identification and antimicrobial resistance detection of sexually transmitted infections

2020 ◽  
Author(s):  
Liqing Zhou ◽  
Andrea Lopez Rodas ◽  
Luz Marina Llangarí ◽  
Natalia Romero Sandoval ◽  
Philip Cooper ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Sexually Transmitted Infections ◽  
Real Time ◽  
Real Time Pcr ◽  
Single Gene ◽  
Vaginal Swab ◽  
Nanopore Sequencing ◽  
Sexually Transmitted ◽  
Negative Controls ◽  
Simultaneous Identification

AbstractObjectivesTo develop a simple DNA sequencing test for simultaneous identification and antimicrobial resistance (AMR) detection of multiple sexually transmitted infections (STIs).MethodsA total of 200 vulvo-vaginal swab samples from 200 female sex workers in Ecuador were initially tested by real-time PCR for Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT), Mycoplasma genitalium (MG) and Trichomonas vaginalis (TV). Samples positive for these STIs and controls were subjected to single gene targeted PCR MinION-nanopore sequencing by using the smartphone operated MinIT.ResultsAmong 200 vulvo-vaginal swab samples 43 were positive for at least one of the STIs by real-time PCR. Single gene targeted nanopore sequencing generally yielded higher pathogen specific reads in positive samples than negative controls. Of the 26 CT, NG or MG infections identified by real-time PCR, 25 were clearly distinguishable from the negative controls using sequence read count. Discrimination of TV positives from controls was poorer as many had low pathogen loads (cycle threshold > 35) with associated low sequence read counts. AMR analysis workflow indicated that 11% of the classified reads aligned with antibiotic resistance genes, all of which identified fluoroquinolone resistance in NG.ConclusionsSingle gene targeted nanopore sequencing for diagnosing and simultaneously identifying key antimicrobial resistance markers for four common STIs, associated with genital tract discharge in women shows promise. Further work to optimise accuracy, reduce costs and improve speed may allow sustainable approaches for managing STIs and emerging AMR in resource poor and laboratory limited settings.

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