Improving molecular tools for global surveillance of measles virus⋆

Details

• Alternative Title:
  J Clin Virol
• Personal Author:
  Bankamp, Bettina ; Byrd-Leotis, Lauren A. ; Lopareva, Elena N. ; Woo, Gibson K.S. ; Liu, Chunyu ; Jee, Youngmee ; Ahmed, Hinda ; Lim, Wilina W. ; Ramamurty, Nalini ; Mulders, Mick N. ; Featherstone, David ; Bellini, William J. ; Rota, Paul A.
• Description:
  Background
  
  The genetic characterization of wild-type measles viruses plays an important role in the description of viral transmission pathways and the verification of measles elimination. The 450 nucleotides that encode the carboxyl-terminus of the nucleoprotein (N-450) are routinely sequenced for genotype analysis.
  
  Objectives
  
  The objectives of this study were to develop improved primers and controls for RT-PCR reactions used for genotyping of measles samples and to develop a method to provide a convenient, safe, and inexpensive means to distribute measles RNA for RT-PCR assays and practice panels.
  
  Study design
  
  A newly designed, genetically defined synthetic RNA and RNA isolated from cells infected with currently circulating genotypes were used to compare the sensitivity of primer pairs in RT-PCR and nested PCR. FTA® cards loaded with lysates of measles infected cells were tested for their ability to preserve viral RNA and destroy virus infectivity.
  
  Results
  
  A new primer pair, MeV216/MeV214, was able to amplify N-450 from viruses representing 10 currently circulating genotypes and a genotype A vaccine strain and demonstrated 100-fold increased sensitivity compared to the previously used primer set. A nested PCR assay further increased the sensitivity of detection from patient samples. A synthetic positive control RNA was developed that produced PCR products that are distinguishable by size from PCR products amplified from clinical samples. FTA® cards completely inactivated measles virus and stabilized RNA for at least six months.
  
  Conclusions
  
  These improved molecular tools will advance molecular characterization of circulating measles viruses globally and provide enhanced quality control measures.
  
  
• Subjects:
  Article DNA Primers Epidemiological Monitoring FTA® Cards Genotype Genotyping Global Health Humans Measles Measles Virus Molecular Diagnostics Molecular Diagnostic Techniques Molecular Epidemiology Reference Standards Reverse Transcriptase Polymerase Chain Reaction RNA, Viral Sensitivity And Specificity

  More +
• Source:
  J Clin Virol. 58(1):176-182.
• Pubmed ID:
  23806666
• Pubmed Central ID:
  PMC5751696
• Document Type:
  Journal Article
• Funding:
  CC999999/Intramural CDC HHS/United States
• Volume:
  58
• Issue:
  1
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:b118e8dd4bb814845cb021b896661f4d7824ecbaee05f011e2ba292dd5a43e93
• Download URL:
  https://stacks.cdc.gov/view/cdc/50738/cdc_50738_DS1.pdf
• File Type:
  [PDF - 1.04 MB ]