Comparison between serum and saliva for the detection of hepatitis A virus RNA

J Virol Methods. 2008 Mar;148(1-2):74-80. doi: 10.1016/j.jviromet.2007.10.020. Epub 2007 Dec 21.

Abstract

Due to the ease of collection, oral fluid is being investigated as an alternative to serum for diagnostic and epidemiological purposes. However, for prospective studies involving hepatitis A virus (HAV) RNA detection, a standard methodology must be developed. In the present study, nested RT-PCR and real-time PCR were optimized and evaluated for HAV detection and quantification, using oral fluid from healthy volunteers (n=20) and paired serum/oral fluid samples from individuals involved in a hepatitis A outbreak (n=78). Using nested RT-PCR, HAV RNA was detected in 50% of oral fluid and in 42% of serum samples from acute cases, as well as in 12% of all samples from cases without IgM and total anti-HAV. Using real-time PCR, HAV RNA was detected in 61% of oral fluid and in 71% of serum samples from acute cases, as well as in 17 and 12%, respectively, from patients without HAV markers. Mean viral loads were 1.7+/-3.24 x 10(3)copies/ml in oral fluid and 2.8+/-6.46 x 10(3)copies/ml in serum. Although nested RT-PCR and real-time PCR both detected HAV RNA in oral fluid, real-time PCR was more sensitive. Oral fluid sample testing could be used as a noninvasive method of detecting HAV RNA during HAV outbreaks.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antibodies, Viral / blood
  • Hepatitis A / diagnosis*
  • Hepatitis A / virology
  • Hepatitis A virus / genetics
  • Hepatitis A virus / isolation & purification*
  • Humans
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / standards
  • Saliva / virology*
  • Sensitivity and Specificity
  • Serum / virology*
  • Viral Load

Substances

  • Antibodies, Viral
  • RNA, Viral