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An integrated cell absorption process and quantitative PCR assay for the detection of the infectious virus in water

Xuan Guo1, Shang Wang2, Chong-Lin Zhao1

  • 1Research Institute of Chemical Defense, No. 1 Huaiyin Road, Beijing 102205, China; State Key Lab of NBC Protection for Civilian, No. 1 Huaiyin Road, Beijing 102205, China.

The Science of the Total Environment
|May 2, 2018
PubMed

Insights

A new integrated cell absorption process and quantitative (reverse transcription) polymerase chain reaction (ICAP-q(RT)PCR) assay offers a highly sensitive and rapid method for detecting infectious viruses in water. This advanced technique improves accuracy and reduces testing time compared to traditional methods.

Area of Science:

  • Environmental microbiology
  • Molecular virology
  • Public health diagnostics

Background:

  • Accurate detection of infectious viruses in water is crucial for public health.
  • Traditional methods like plaque assays and direct qPCR have limitations in sensitivity and speed.
  • Existing integrated cell culture (ICC)-qPCR methods offer improvements but can be time-consuming.

Purpose of the Study:

  • To develop and validate a novel assay for sensitive and rapid detection of infectious viruses.
  • To improve upon existing molecular diagnostic tools for water quality monitoring.
  • To assess the prevalence of specific viruses in environmental water samples.

Main Methods:

  • Development of an integrated cell absorption process and quantitative (reverse transcription) polymerase chain reaction (ICAP-q(RT)PCR) assay.
  • Detection of viral nucleic acid (RNA or DNA) during early stages of viral attachment and entry.
  • Validation using poliovirus, adenovirus, rotavirus, and astrovirus in water samples.

Main Results:

  • ICAP-q(RT)PCR detected as low as 0.2 TCID50/mL of poliovirus or adenovirus after 4h incubation.
  • The assay demonstrated significantly higher sensitivity than plaque assays.
  • Consistent results with ICC-qPCR were achieved in a shorter timeframe for field samples.
  • Detected poliovirus in 50% of river water samples, aligning with ICC-qPCR, but differing from plaque assay and direct qPCR results.

Conclusions:

  • The developed ICAP-q(RT)PCR method significantly reduces test duration and enhances sensitivity for infectious viruses.
  • This assay provides an effective and quantitative tool for detecting infectious viruses in water environments.
  • ICAP-q(RT)PCR offers improved accuracy compared to direct qPCR and plaque assays for environmental water monitoring.
Keywords:
Cell absorption based processEnvironmental samplesInfectious virus detectionQuantitative polymerase chain reaction

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