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Background <p>Adverse pregnancy outcomes (APOs), including spontaneous abortion, stillbirth, and neonatal sepsis, are major contributors to maternal and neonatal morbidity and mortality worldwide, particularly in low- and middle-income countries. Infectious agents are increasingly recognized as key, yet underdiagnosed, etiological factors in APOs. The study was undertaken with the following objectives: (1) To develop and validate in-house real-time PCR assay to detect a broad spectrum of infectious agents beyond standard TORCH pathogens. (2) To conduct cross-sectional molecular surveillance to assess the prevalence of pathogens of interest in patients with adverse pregnancy outcomes and neonatal sepsis at a tertiary care hospital in Central India.</p> Methods <p>This cross-sectional study was conducted at two tertiary care centres in Bhopal, India. Clinical samples (blood, urine, placental tissue, Cerebrospinal fluid) were collected from women with cases of Adverse Pregnancy Outcome. Real-time PCR assays targeting <i>Brucella spp</i>., Human Parvovirus B19, Enterovirus, Cytomegalovirus, Epstein-Barr Virus, Hepatitis E Virus, <i>Listeria monocytogenes</i>, and other pathogens were developed and validated against commercial CE-IVD kits.</p> Results <p>A total of 99 cases of adverse pregnancy outcomes and 27 neonatal sepsis cases were analysed. The assay demonstrated 90-99.3% amplification efficiency and a detection limit of 1–6 copies/reaction. Pathogen detection rates ranged from 44.2% to 60.3%. <i>Brucella spp</i>. (12-19.2%), Enterovirus (5.2–15.6%) was the most prevalent infection. Neonatal infections were detected in 44.4% of neonatal sepsis cases, with co-infections observed in 14.8% of affected neonates.</p> Conclusion <p>The study underscores the burden of underrecognized pathogens in adverse pregnancy outcomes and neonatal sepsis, highlighting the diagnostic value of molecular testing in prenatal and neonatal care. Integration of broader infectious screening into routine antenatal care may facilitate earlier interventions and improve outcomes in endemic settings.</p> Trial registration <p>Not applicable.</p>

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Development of in-house PCR assays and cross-sectional molecular surveillance of infectious pathogens in adverse pregnancy outcomes and neonatal sepsis in central India

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年2月5日

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  • Kashmi Sharma,
  • Rohan Shrivastava,
  • Somesh Mishra,
  • Debasis Biswas,
  • K. Pushpalatha,
  • Ashwin A. Raut,
  • Jitendra Singh,
  • Rupinder Kaur Kanwar,
  • Shashank Purwar,
  • Ankur Joshi,
  • Shashwati Nema,
  • Shweta Patel,
  • Jyotsana Shrivastava,
  • Aruna Kumar,
  • Shabana Sultan,
  • Manjusha Goel,
  • Pallavi Singh,
  • Poorva Gohiya,
  • Ram K. Nema,
  • Megha Katare Pandey

摘要

Background

Adverse pregnancy outcomes (APOs), including spontaneous abortion, stillbirth, and neonatal sepsis, are major contributors to maternal and neonatal morbidity and mortality worldwide, particularly in low- and middle-income countries. Infectious agents are increasingly recognized as key, yet underdiagnosed, etiological factors in APOs. The study was undertaken with the following objectives: (1) To develop and validate in-house real-time PCR assay to detect a broad spectrum of infectious agents beyond standard TORCH pathogens. (2) To conduct cross-sectional molecular surveillance to assess the prevalence of pathogens of interest in patients with adverse pregnancy outcomes and neonatal sepsis at a tertiary care hospital in Central India.

Methods

This cross-sectional study was conducted at two tertiary care centres in Bhopal, India. Clinical samples (blood, urine, placental tissue, Cerebrospinal fluid) were collected from women with cases of Adverse Pregnancy Outcome. Real-time PCR assays targeting Brucella spp., Human Parvovirus B19, Enterovirus, Cytomegalovirus, Epstein-Barr Virus, Hepatitis E Virus, Listeria monocytogenes, and other pathogens were developed and validated against commercial CE-IVD kits.

Results

A total of 99 cases of adverse pregnancy outcomes and 27 neonatal sepsis cases were analysed. The assay demonstrated 90-99.3% amplification efficiency and a detection limit of 1–6 copies/reaction. Pathogen detection rates ranged from 44.2% to 60.3%. Brucella spp. (12-19.2%), Enterovirus (5.2–15.6%) was the most prevalent infection. Neonatal infections were detected in 44.4% of neonatal sepsis cases, with co-infections observed in 14.8% of affected neonates.

Conclusion

The study underscores the burden of underrecognized pathogens in adverse pregnancy outcomes and neonatal sepsis, highlighting the diagnostic value of molecular testing in prenatal and neonatal care. Integration of broader infectious screening into routine antenatal care may facilitate earlier interventions and improve outcomes in endemic settings.

Trial registration

Not applicable.