Rapid detection of measles virus using reverse transcription loop-mediated isothermal amplification coupled with a disposable lateral flow device

Diagn Microbiol Infect Dis. 2016 Jun;85(2):168-73. doi: 10.1016/j.diagmicrobio.2016.02.023. Epub 2016 Mar 3.

Abstract

The measles virus (MeV) causes a highly contagious disease and efforts to reduce its spread are critical. A reverse transcription loop-mediated isothermal amplification assay coupled with a disposable lateral flow device (RT-LAMP-LFD) was developed for the rapid detection of MeV. The assay was performed in 40 min at an optimal temperature of 58 °C, with endpoint results visualized directly. A probe that was complementary to the RT-LAMP amplicon was designed to enhance assay specificity. Detection limit of the assay was 8.8 copies/μL synthetic RNA, which equals the sensitivity of real-time RT-PCR. Clinical specimens were used to validate the RT-LAMP-LFD in provincial Center for Disease Control and Prevention (CDC) (n = 245) and six municipal CDCs (n = 249). The results obtained using RT-LAMP-LFD and real-time RT-PCR were highly concordant. The RT-LAMP-LFD is rapid, stable, and does not require expensive equipment, which can be used for routine MeV monitoring in CDC laboratories.

Keywords: Lateral flow device; Measles virus; Rapid detection; Reverse transcription loop-mediated isothermal amplification.

Publication types

  • Evaluation Study

MeSH terms

  • Humans
  • Measles / diagnosis*
  • Measles virus / genetics
  • Measles virus / isolation & purification*
  • Nucleic Acid Amplification Techniques / instrumentation*
  • Nucleic Acid Amplification Techniques / methods*
  • Oligonucleotide Probes / genetics
  • Sensitivity and Specificity
  • Temperature
  • Time Factors

Substances

  • Oligonucleotide Probes