Enhancing enteric pathogen detection: implementation and impact of multiplex PCR for improved diagnosis and surveillance.

BACKGROUND: Syndromic surveillance of acute gastroenteritis plays a significant role in the diagnosis and management of gastrointestinal infections that are responsible for a substantial number of deaths globally, especially in developing countries. In Lebanon, there is a lack of national surveillance for acute gastroenteritis, and limited data exists regarding the prevalence of pathogens causing diarrhea. The one-year study aims to investigate the epidemiology of common gastrointestinal pathogens and compare our findings with causative agents of diarrhea reported by our study collaborative centers. METHODS: A multicenter, cross-sectional study was conducted over a one-year period. A total of 271 samples were obtained from outpatients and inpatients presenting with symptoms of acute gastroenteritis at various healthcare facilities. The samples were then analyzed using Allplex gastrointestinal assay that identifies a panel of enteric pathogens. RESULTS: Overall, enteropathogens were detected in 71% of the enrolled cases, 46% of those were identified in patients as single and 54% as mixed infections. Bacteria were observed in 48%, parasites in 12% and viruses in 11%. Bacterial infections were the most prevalent in all age groups. Enteroaggregative E. coli (26.5%), Enterotoxigenic E. coli (23.2%) and Enteropathogenic E. coli (20.3%) were the most frequently identified followed by Blastocystis hominis (15.5%) and Rotavirus (7.7%). Highest hospitalization rate occurred with rotavirus (63%), Enterotoxigenic E. coli (50%), Blastocystis hominis (45%) and Enteropathogenic E. coli (43%). Enteric pathogens were prevalent during summer, fall and winter seasons. CONCLUSIONS: The adoption of multiplex real-time PCR assays in the diagnosis of gastrointestinal infections has identified gaps and improved the rates of detection for multiple pathogens. Our findings highlight the importance of conducting comprehensive surveillance to monitor enteric infections. The implementation of a syndromic testing panel can therefore provide healthcare professionals with timely and accurate information for more effective treatment and public health interventions.

Comparison of two commercial multiplex PCR assays for the detection of sexually transmitted infections.

INTRODUCTION: Multiplex molecular panels are replacing conventional methods for the detection of sexually transmitted infections. In the current study, we evaluated the performance of two commercial multiplex assays, EUROArray STI and Allplex STI essential assays, for detecting six sexually transmitted infections. METHODOLOGY: The diagnostic performance of the EUROArray STI and Allplex STI essential assays was evaluated against a panel of 105 positive DNA samples identified by in-house real-time PCR assays including Ureaplasma parvum, Ureaplasma urealyticum, Mycoplasma hominis, Trichomonas vaginalis, Chlamydia trachomatis, and Neisseria gonorrhea. Samples from healthy subjects, negative for any microorganism, were used as negative controls. RESULTS: Of the 105 positive specimens, 103 (98%) were tested positive by Allplex and 102 (97%) by EUROArray. Among the 51 negative samples that were tested by in house assay, 48 (94%) were tested negative by Allplex assay and 43 (84%) by EUROArray assay. The overall sensitivity of EUROArray and Allplex were 97.1% and 98.1% with an accuracy of 92.9% and 96.7%, respectively. The overall assay specificity was 94.1% for Allplex assay and 84.3% for EUROArray assay, The sensitivity of both kits to all targeted microorganisms ranged from 55.6% to 100%, with the lowest sensitivity noted for Trichomonas vaginalis. CONCLUSIONS: Diagnostic performance varies depending on the method used to detect the targeted pathogens, the assay manipulation, and the cost. This study showed sensitivity, specificity, and accuracy characteristics for two kits commonly used to detect STIs, which will guide the choice for an appropriate multiplex PCR platform.