Clinical Signs and Symptoms in Sexually Transmitted Infections Confirmed by Multiplex PCR: Practical Tips for Clinicians

Background Sexually transmitted infections (STIs) refer to infections transmitted through sexual contact, including asymptomatic cases. Data on co-infections among STI pathogens based on different specimen types have been accumulating with the increasing use of multiplex PCR for STI diagnosis. This study retrospectively analyzed the epidemiological characteristics of STIs using multiplex PCR test results from South Korea(2018-2024), focusing on co-infections. Methods We analyzed multiplex PCR test results for 12 STI pathogens from January 2018 to September 2024 in individuals aged 10 to 90, conducted by Eone Laboratories. The pathogens included Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT), Mycoplasma hominis (MH), Trichomonas vaginalis (TV), Ureaplasma urealyticum (UU), Mycoplasma genitalium (MG), Herpes simplex virus type 1 (HSV1), Herpes simplex virus type 2 (HSV2), Gardnerella vaginalis (GV), Treponema pallidum (TP), Candida albicans (CA), and Ureaplasma parvum (UP). The dataset included information on gender, age, specimen type, and test date. Multiplex PCR reaction was performed using NextGene STI-12 Detection Kit (EONEBIOTECH, Republic of Korea) with an CFX 96 real-time PCR system(Bio-Rad, Republic of Korea). We analyzed the positivity rates by gender, age, and specimen type. Additionally, association rule mining was applied to evaluate co-infection patterns, and lift indices were calculated to assess the strength of associations between pathogens. Result From 2,694,547 specimens, 32,334,564 STI test results were obtained, with 3,093,258 positive cases (9.6%). Female specimens comprised 70.1%, and male 29.9%.. Among the specimens, 40.0% were negative for all pathogens, 25.8% were positive for a single pathogen, and 34.2% had co-infections with two or more pathogens. Pathogen positivity rates showed that in females, GV, UP, CA, UU, and MH had a positivity rate above 10%, while in males, UU, CT, and UP had a positivity rate exceeding 5%. NG and CT had higher positivity in males than females, peaking in individuals in their 10s and 20s, and declining with increasing age. Age-specific positivity rates revealed that, exception of TV and GV in males and HSV2 in females, all STI pathogens peaked in individuals in their 10s and 20s. In females, UU, MH, TV, GV, and UP exhibited a biphasic peak pattern, with the first peak occurring in individuals in their 10s and 20s, followed by a second peak in their 50s and 60s. A comparison of positivity rates by specimen type in female samples revealed that vaginal discharge specimens had 1.28 to 8.19 times higher positivity rates than urine specimens, indicating higher sensitivity for STI detection in female. Co-infection analysis revealed that CT-NG and TV-MH had a lift index of =3 in both males and females, indicating a strong co-occurrence. When lift indices were analyzed separately for individuals aged 10–30 and those over 30, the absolute values varied by age group; however, co-infection patterns were consistent within the same gender but differed between males and females. Conclusion This study assessed the associations among major STI pathogens based on multiplex PCR test results and analyzed their epidemiological characteristics. Further research is needed to incorporate clinical symptom data and elucidate the biological interactions of highly associated pathogens.

Performance of Anyplex™ II multiplex real-time PCR for the diagnosis of seven sexually transmitted infections: comparison with currently available methods

BackgroundThe burden of sexually transmitted infections (STIs) has been increasing in Kenya, as is the case elsewhere in sub-Saharan Africa, while measures for control and prevention are weak. The objectives of this study were to (1) describe healthcare provider (HCP) knowledge and practices, (2) explore HCP attitudes and beliefs, (3) identify structural and environmental factors affecting STI management, and (4) seek recommendations to improve the STI program in Kenya.Methods and findingsUsing individual in-depth interviews (IDIs), data were obtained from 87 HCPs working in 21 high-volume comprehensive HIV care centers (CCCs) in 7 of Kenya’s 8 regions. Transcript coding was performed through an inductive and iterative process, and the data were analyzed using NVivo 10.0. Overall, HCPs were knowledgeable about STIs, saw STIs as a priority, reported high STI co-infection amongst people living with HIV (PLHIV), and believed STIs in PLHIV facilitate HIV transmission. Most used the syndromic approach for STI management. Condoms and counseling were available in most of the clinics. HCPs believed that having an STI increased stigma in the community, that there was STI antimicrobial drug resistance, and that STIs were not prioritized by the authorities. HCPs had positive attitudes toward managing STIs, but were uncomfortable discussing sexual issues with patients in general, and profoundly for anal sex. The main barriers to the management of STIs reported were low commitment by higher levels of management, few recent STI-focused trainings, high stigma and low community participation, and STI drug stock-outs. Solutions recommended by HCPs included formulation of new STI policies that would increase access, availability, and quality of STI services; integrated STI/HIV management; improved STI training; increased supervision; standardized reporting; and community involvement in STI prevention. The key limitations of our study were that (1) participant experience and how much of their workload was devoted to managing STIs was not considered, (2) some responses may have been subject to recall and social desirability bias, and (3) patients or clients of STI services were not interviewed, and therefore their inputs were not obtained. While considering these limitations, the number and variety of facilities sampled, the mix of staff cadres interviewed, the use of a standardized instrument, and the consistency of responses add strength to our findings.ConclusionsThis study showed that HCPs understood the challenges of, and solutions for, improving the management of STIs in Kenya. Commitment by higher management, training in the management of STIs, measures for reducing stigma, and introducing new policies of STI management should be considered by health authorities in Kenya.

Parapneumonic effusion in children: Rapid pathogen detection in pleural fluid using multiplex bacterial PCR.

To determine the etiology of genital ulcer disease (GUD) among patients attending sexually transmitted disease (STD) clinics in Pune, India, and to examine the relationship to HIV infection and compare the clinical diagnosis of GUD with the results of a multiplex polymerase chain reaction (M-PCR) assay for Treponema pallidum, herpes simplex virus (HSV), and Hemophilus ducreyi infection. Between June 20, 1994, and September 26, 1994, 302 patients with a genital ulcer were evaluated. Clinical etiology of GUD was based on physical appearance and microbiologic evaluations which included darkfield microscopy and serology for syphilis. Swabs of each genital ulcer were tested for HSV antigen by enzyme immunoassay (Herpchek; Dupont, Wilmington, DE) and processed in a multiplex PCR assay (M-PCR; Roche, Branchburg, NJ) for simultaneous detection of HSV, Treponema pallidum, and Hemophilus ducreyi. Two hundred seventy-seven men and 25 women with a median age of 25 were evaluated. The seroprevalence of HIV was 22.2%. The etiology of GUD as determined by M-PCR was HSV (26%), H. ducreyi (23%), T. pallidum (10%), and multiple infections (7%); no etiology was identified in 34%. HIV seroprevalence was higher among those patients positive for HSV compared with other etiologies (OR = 2.1, CI: 1.2-3.7; p = 0.01). When compared with M-PCR, the Herpchek test was 68.5% sensitive and 99.5% specific. Darkfield detection for T. pallidum was 39% sensitive and 82% specific, in contrast to rapid plasma reagin and fluorescent treponemal antibody absorption test, which was 66% sensitive and 90% specific. Clinical diagnosis alone or in combination with basic laboratory tests showed poor agreement with M-PCR.

This study primarily aimed to investigate the prevalence of human papillomavirus (HPV) and other common pathogens of sexually transmitted infections (STIs) in spermatozoa of infertile men and their effects on semen parameters. These pathogens included Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Mycoplasma genitalium , herpes simplex virus 2, Neisseria gonorrhoeae, Enterococcus faecalis, Streptococcus agalactiae, Pseudomonas aeruginosa , and Staphylococcus aureus . A total of 1951 men of infertile couples were recruited between 23 March 2023, and 17 May 2023, at the Department of Reproductive Medicine of The First People's Hospital of Yunnan Province (Kunming, China). Multiplex polymerase chain reaction and capillary electrophoresis were used for HPV genotyping. Polymerase chain reaction and electrophoresis were also used to detect the presence of other STIs. The overall prevalence of HPV infection was 12.4%. The top five prevalent HPV subtypes were types 56, 52, 43, 16, and 53 among those tested positive for HPV. Other common infections with high prevalence rates were Ureaplasma urealyticum (28.3%), Ureaplasma parvum (20.4%), and Enterococcus faecalis (9.5%). The prevalence rates of HPV coinfection with Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Mycoplasma genitalium , herpes simplex virus 2, Neisseria gonorrhoeae, Enterococcus faecalis, Streptococcus agalactiae , and Staphylococcus aureus were 24.8%, 25.4%, 10.6%, 6.4%, 2.4%, 7.9%, 5.9%, 0.9%, and 1.3%, respectively. The semen volume and total sperm count were greatly decreased by HPV infection alone. Coinfection with HPV and Ureaplasma urealyticum significantly reduced sperm motility and viability. Our study shows that coinfection with STIs is highly prevalent in the semen of infertile men and that coinfection with pathogens can seriously affect semen parameters, emphasizing the necessity of semen screening for STIs.

The World Health Organization's Global strategy for the prevention and control of sexually transmitted infections (STIs): 2006-2015 highlighted the need for STI surveillance as a cornerstone for national programmes. Yet, in many countries of the world, little or no surveillance exists and, when it does, it is often clinical in nature. Much of the world's resource-poor areas use the syndromic management approach, which includes a recommendation for periodic surveillance of antimicrobial resistance in Neisseria gonorrhoeae. It is also important to perform aetiological surveillance, to assess the common causes of the main STI syndromes, such as genital ulceration (GUS), male urethritis syndrome (MUS) and the vaginal discharge syndrome (VDS). This allows observation of trends and ensures that the drugs used in the syndromic management flow chart as still valid. South Africa started to build a national microbiological and clinical surveillance programme in 2004. Prior to that, microbiological data came from surveillance among particular core groups, such as miners, that could not be extrapolated to the general population. 30 sentinel sites (primary healthcare facilities) were set up in each of the country's nine provinces for the purpose of enhanced clinical surveillance. Data were collected on all the main syndromes in terms of episodes per year. At the same time, microbiological surveillance was initiated in the following provinces: the Northern Cape, Mpumalanga, the Western Cape and Gauteng. Plans are to conduct further surveillance in the Free State and possibly the Eastern Cape later in 2007. Within each province, one primary health care facility was chosen on the criteria of a large STO caseload and proximity to the laboratory doing the initial culturing of N. gonorrhoeae. Consecutive patients were recruited using informed consent and anonymous specimens collected. Patients were treated syndromically in the normal manner according to national STI management guidelines. Gonococcal isolates, obtained from men with urethral discharge, were tested for ciprofloxacin and ceftriaxone resistance using E tests. In addition, swabs were collected from MUS patients and VDS patients for multiplex polymerase chain reaction (M-PCR) based testing for the following four pathogens: N. gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis and Mycoplasma genitalium. Ulcer swabs were also tested by M-PCR for herpes simplex virus (HSV), Haemophilus ducreyi and Treponema pallidum. A separate PCR was used to test the extracted DNA for C. trachomatis L1-L3. Serum was taken from all participants and tested for syphilis (RPR plus TPPA), HSV-2 and HIV antibodies. Key findings have confirmed the decline of chancroid to below 1% of genital ulcers and the predominance of genital herpes as the major cause of genital ulceration in South Africa. Gonorrhoea continues to be the major cause of urethritis in men and prevalence far exceeds Chlamydial infection. Approximately 10% of men with MUS are also infected/colonized with T. vaginalis. Only about one third of VDS cases appear to be caused by sexually transmitted pathogens. HIV infection rates exceed those recorded in the annual antenatal surveys and are highest among genital ulcer patients (70%). RPR seropositivity in non-ulcer patients is around 5% and antibodies to HSV-2 occur in about 50!!60% of patients overall. The surveillance has also demonstrated alarming rises in the prevalence of ciprofloxacin resistant gonorrhoea since 2004.

Background: Bacterial vaginitis (BV) and Trichomonas vaginitis are the most frequently recurring infectious diseases in women. Therefore, accurate tests for post-treatment follow-up are required. A multiplex PCR assay allows for the simultaneous detection of multiple pathogens in a single specimen. In this study, we assessed the clinical implications of multiplex PCR detection of fastidious microorganisms causing vaginitis. Methods: A total of 216 vaginitis patients who presented to Chung-Ang University Yongsan Hospital with more than one positive result on multiplex PCR (Trichomonas vaginalis (TV), Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT), Ureaplasma urealyticum (UU), Mycoplasma genitalium (MG), Mycoplasma hominis (MH)) were retrospectively enrolled in this study. Each patient’s clinical symptoms, initial treatment and follow-up for BV, and other related test results were also retrospectively reviewed. Results: The most commonly reported symptom was abnormal discharge, followed by pruritis (73.1%), lower abdominal pain (38.4%), urination difficulties (13%), and others such as fever. According to the multiplex PCR results, there were 116 cases (35.8%) of MH, 86 cases (26.5%) of UU, 62 cases (19.1%) of CT, and 84 cases (38.9%) were mixed infections. Among those patients with single infections, treatment changed for 63 cases (65.6%) while treatment remained unchanged for 17 (17.7%) after PCR results were reported. Conclusion: The diagnosis of BV using multiplex PCR is clinically effective and the results of which can be incorporated in antibiotic selection for patients with multiple sexually transmitted diseases (STD). Multiplex PCR may be especially helpful in the diagnosis of patients in whom the differentiation of STD pathogens is difficult using traditional methods. (Korean J Clin Microbiol 2011;14:30-35)

"Sex in the Time of COVID": Clinical Guidelines for Sexually Transmitted Disease Management in an Era of Social Distancing.

BackgroundDelays in the reporting of STD testing sometimes result in inappropriate patient care where a patient must be called back in for treatment. Some STDs may be missed because not...

Population-based interventions for reducing sexually transmitted infections, including HIV infection.

Sexually transmitted infections (STIs) cause considerable morbidity worldwide and, depending on the specific pathogen, may lead to serious complications in the female reproductive tract. Incarcerated women are particularly vulnerable to health problems with a disproportionate high rate of STIs, including infections with human papillomavirus (HPV). Here, cervical swab samples collected from 299 women (18 to 64 years) living in one of the women's prisons of São Paulo, Brazil were submitted for liquid-based cytology to determine the prevalence of precancerous lesions. Furthermore, direct detection of 30 genital HPV genotypes (18 high-risk and 12 low-risk types) and 11 additional STIs (Chlamydia trachomatis, Neisseria gonorrhoeae, Herpes simplex virus 1 and 2, Haemophilus ducreyi, Mycoplasma genitalium and hominis, Treponema pallidum, Trichomonas vaginalis, Ureaplasma parvum and urealyticum) were performed by molecular typing using two PCR-based DNA microarray systems, i.e., EUROArray HPV and EUROArray STI (EUROIMMUN), respectively. The overall prevalence of cytological abnormalities was 5.8%, including five women with low-grade and five women with high-grade squamous intraepithelial lesions. The overall prevalence of HPV was 62.2, and 87.1% of the HPV-positive women were infected with oncogenic high-risk (HR) HPV types. HPV types 16 (24.1%), 33 and 52 (both 10.4%) were the most frequently detected. The prevalence of the other STIs was 72.8%. Up to four different pathogens were found in the infected women, the most frequent being Ureaplasma parvum (45.3%), Mycoplasma hominis (36.2%) and Trichomonas vaginalis (24.8%). The high number of HR-HPV infections and other STIs described here highlights the fact that the Brazilian female prison population requires more attention in the country's health policies. The implementation of screening programs and treatment measures might contribute to a decrease in the incidence of STIs and cervical cancer in this vulnerable population. However, for such measures to be effective, further studies are needed to investigate the best practice to get more women to engage in in-prison prevention programs, e.g., through offering further sexual health education and self-sampling.

Background: Sexually transmitted diseases (STD) are a major cause of infertility, long-term disability, ectopic pregnancy, and premature birth. Therefore, the development of fast and low-cost laboratory STD diagnostic screening methods will contribute to reducing STD-induced reproductive tract damage and improve women's health worldwide. In this study, we evaluated a novel multiplex real-time PCR melting curve assay method for the simultaneous detection of 9 STD pathogens, including Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, Mycoplasma hominis, Ureaplasma urealyticum, Ureaplasma parvum, and herpes simplex virus.Methods: The analytical performance of the method, including its limit of detection (LOD), specificity, repeatability, and effect on different DNA extraction kits were evaluated. Additionally, we obtained 1,328 clinical specimens from 3 hospitals to detect the 9 STD pathogens using multiplex real-time PCR melting curve and Sanger sequencing, to evaluate the sensitivity, specificity, and consistency of the assay method.Results: The results showed that the analytical sensitivity of the novel multiplex real-time PCR melting curve assay is very excellent, with LOD of DNA corresponding to <200 copies/μL for the DNA of the 9 STDs and 1.00 × 104 color change unit /ml for those of UU and UP. Additionally, this assay demonstrated excellent analytical specificity, excellent repeatability, and its results had no effect of different DNA extraction kits. The performance, in terms of sensitivity (91.06–100%) and specificity (99.14–100%), was remarkable, since the consistency between it and Sanger sequencing was more than 0.85 in the clinic.Conclusion: The novel multiplex real-time PCR melting curve assay method has high sensitivity and specificity, relatively low cost, and simple to use for the simultaneous detection of 9 STD pathogens in genitourinary secretions.

AAUS guidelines 2021 revision sexually transmitted infection (STIs) diagnostic strategy for STI

Chapter 6 - Sexually transmitted viral infections