[Use of multiplex PCR method with real-time detection for differential diagnosis of respiratory viral infections]

Zh Mikrobiol Epidemiol Immunobiol. 2009 Jan-Feb:(1):67-70.
[Article in Russian]

Abstract

Multiplex real-time polymerase chain reaction test-system with fluorescent detection (RT-PCR) for simultaneous identification of main agents of acute respiratory viral infections: influenza A (IAV) and B viruses (IBV), parainfluenza viruses types 1, 2, 3, 4 (PIV 1 - 4), adenoviruses (ADV), respiratory syncitial virus (RSV), rhinoviruses (RV) and enteroviruses (EV), in presence internal positive control (IPC) represented by vaccine strain of rubella virus RA 27/3. Using multiplex RT-PCR method, respiratory viruses were detected in 116 out of 226 clinical samples (nasal swabs) obtained from patients with symptoms of acute respiratory infection: in 68 (58.6%) samples--IBV; in 21 (18.1%)--IAV; in 12 (10.3%) --RV; in 6 (5.2%)--PIV 2; in 4--(3.4%) ADV; in 3 (2.6%)--RSV; in 2 (1.7%)--EV; in 2 (1.7%)--PIV 4; in 1 (0.9%)--PIV 3; in 1 (0.9%)--PIV 1. Mixed infection was observed in 4 (3.4%) patients. PCR assay allowed to reveal various respiratory viruses in 51.3% of samples. At the same time samples were tested for the presence of 12 respiratory viruses--IAV, IBV, PIV 1 - 4, RSV, RV, metapneumoviruses, and coronaviruses NL63, 229E and OC43--in the presence of IPC represented by equine arteritis virus using analogous PCR test-system provided by medical center of Leiden university. Results of tests for detection of IAV, IBV, RSV, PIV 1 - 4, and RV, analyzed by both systems, agreed in 94%. Multiplex format of RT-PCR performing significantly reduces time and cost of the test, which make it suitable and effective instrument of epidemiological studies.

Publication types

  • Evaluation Study

MeSH terms

  • Acute Disease
  • Adenoviridae / isolation & purification
  • DNA Primers
  • DNA, Viral / analysis
  • Diagnosis, Differential
  • Enterovirus / genetics
  • Enterovirus / isolation & purification
  • Humans
  • Influenza A virus / genetics
  • Influenza A virus / isolation & purification
  • Influenza B virus / genetics
  • Influenza B virus / isolation & purification
  • Nasal Mucosa / virology
  • Paramyxovirinae / genetics
  • Paramyxovirinae / isolation & purification
  • Polymerase Chain Reaction / methods*
  • RNA, Viral / analysis
  • Respiratory Syncytial Viruses / genetics
  • Respiratory Syncytial Viruses / isolation & purification
  • Respiratory Tract Infections / diagnosis*
  • Respiratory Tract Infections / virology
  • Rhinovirus / genetics
  • Rhinovirus / isolation & purification
  • Sensitivity and Specificity
  • Virus Diseases / diagnosis*

Substances

  • DNA Primers
  • DNA, Viral
  • RNA, Viral