A simple smart amplification assay for the rapid detection of human cytomegalovirus in the urine of neonates

J Virol Methods. 2014 Nov:208:160-5. doi: 10.1016/j.jviromet.2014.07.034. Epub 2014 Aug 7.

Abstract

Human cytomegalovirus (CMV) is the most common cause of infection-related congenital abnormalities in neonates and the leading cause of non-hereditary sensorineural hearing loss in childhood. In addition, the number of low-birth-weight infants has recently increased, especially in Japan, in association with an increasing frequency of postnatal CMV infections transferred through raw breast milk. The increase in the number of congenital CMV and postnatal CMV infections in low-birth-weight infants requires rapid detection at the bedside in order to ensure a correct diagnosis and provide early anti-viral therapy. In this report, a simplified smart amplification (SMAP) method was developed to detect CMV in the urine of neonates. This method does not require DNA extraction, and the DNA amplification procedure is performed under isothermal conditions. Therefore, it takes only 60 min to detect CMV in a urine sample, and CMV DNA was rapidly detectable in symptomatic infants. In brief, this SMAP-based assay provides a simple, rapid and efficient method for detecting human CMV at the bedside.

Keywords: Human cytomegalovirus; Isothermal nucleotide amplification; Real-time PCR; Smart amplification.

MeSH terms

  • Child, Preschool
  • Cytomegalovirus / isolation & purification*
  • Cytomegalovirus Infections / diagnosis*
  • Cytomegalovirus Infections / virology
  • Humans
  • Infant, Newborn
  • Japan
  • Molecular Diagnostic Techniques / methods*
  • Nucleic Acid Amplification Techniques / methods*
  • Time Factors
  • Urine / virology*