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Development of a multiplex qPCR assay for the detection of causative agents for TORCH in neonates

microbiology
molecular-biology
infectious-diseases
pediatrics
obstetrics-and-gynecology

Abstract

Background: The acronym TORCH refers a group of pathogens including Toxoplasma gondii (T. gondii), Treponema pallidum (T. pallidum), Cytomegalovirus (CMV), and Herpes Simplex Viruses (HSV) which cause congenital TORCH infection. Serological tests or viral culture are limited due to cross-reactivity, low accuracy and prolonged turnaround times. Methods: A rapid multiplex quantitative real-time PCR (qPCR) assay was developed to detect congenital TORCH pathogens. Results: The study demonstrated a lower detection limit (LOD) of 10 ag/µL for T. gondii, T. pallidum, and CMV, and 1 ag/µL for HSV-1 and HSV-2. Additionally, the assay showed 100% for analytical specificity against 38 organisms. The PCR efficiency exhibited ranging between 95% and 109% for each organism, with a coefficient of variation (CV) of less than 3%. Conclusion: This multiplex qPCR shows significant potential for improving the speed and accuracy of diagnosing TORCH pathogens in neonates.
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