Clinical and Molecular Diagnostic Profiling of Vaginitis Using Multiplex Real-Time PCR: A Multicenter Study

<b>Background:</b> Vaginal infections often present with overlapping symptoms and involve single or multiple pathogens. However, the relationship between clinical symptoms and molecularly defined vaginal pathogen profiles, especially in multi-pathogen infections, remains poorly characterized in a routine care setting. This study exams the connection between vaginal symptoms and pathogen profiles among women with vaginitis in Northern Vietnam. <b>Methods:</b> We conducted a multicenter cross-sectional study of women with vaginitis at Bac Ninh CDC and Hanoi Obstetrics and Gynecology Hospital between December 2023 and December 2024. Baseline demographics and clinical symptoms were assessed by physicians. Vaginal swabs were collected for pH measurement and pathogen detection using multiplex real-time PCR. The correlation was analyzed using logistic regression in GraphPad Prism v10.1.1. <b>Results:</b> Among 289 symptomatic women, abnormal vaginal discharge and itching were the most common symptoms. <i>Gardnerella vaginalis</i> was the most commonly detected pathogen, occurring alone or in combination with <i>Candida albicans</i>, <i>Mycoplasma hominis</i>, and other genital pathogens. Multi-pathogen infection was associated with abnormal vaginal discharge (OR = 5.44), itching (OR = 2.13), and elevated vaginal pH (OR = 4.70). Women at the tertiary hospital showed greater symptom burden (OR = 1.75) and higher prevalence of multi-pathogen infections (OR = 9.75) than those attending the provincial CDC. <b>Conclusions:</b> Multiplex real-time PCR combined with simple clinical indicators (symptom clustering and vaginal pH) provides practical diagnostic value for identifying multi-pathogen infections in symptomatic women. This integrated approach may support more accurate etiologic diagnosis and guide rational testing strategies, particularly in resource-limited settings.