Measles virus genotyping by nucleotide-specific multiplex PCR

Journal of Clinical Microbiology
Jacques R KremerClaude P Muller

Abstract

A simple genotyping method based on multiplex PCR has been developed to discriminate between all active measles virus (MV) clades and genotypes (A, B3.1, B3.2, C2, D2-D9, G2-G3, and H1-H2). The sequencing reaction was replaced by six multiplex PCRs: one to identify the clade and five to identify the respective genotype. Primers were sensitive to clade- and genotype-specific nucleotides and generated fragments of type-specific sizes that were analyzed by conventional agarose gel electrophoresis. On the basis of all published MV sequences, positive and negative predictive values of 99.2% and 98.6% were calculated. Variability in the primer binding sites, which could potentially reduce sensitivity, was very limited among published sequences. As new genotypes are described, additional specific primers can be included in the multiplex PCR with relative ease. Although sequencing remains the "gold standard," the present method should facilitate MV genotyping especially in developing countries and will therefore contribute to enhanced MV control and elimination strategies as recommended by the World Health Organization.

References

May 9, 2000·Lancet·P A RotaK B Chua
Mar 21, 2001·Vaccine·M N MuldersC P Muller
Jun 14, 2001·Archives of Virology·P R BarreroA S Mistchenko
Jul 4, 2002·Emerging Infectious Diseases·Doris ChiboChristopher Birch
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Dec 17, 2003·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Oliver KirschbergStephan Schaefer

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Citations

Jun 17, 2011·The Journal of Infectious Diseases·Paul A RotaDavid Featherstone
Oct 6, 2006·Journal of Clinical Microbiology·Alexander A NeverovVladimir E Chizhikov
Feb 28, 2013·BMC Infectious Diseases·Vianney TricouIonela Gouandjika-Vasilache
Mar 21, 2008·Journal of Virological Methods·Judith M HübschenClaude P Muller
Aug 22, 2007·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Xuelian YuReena Ghildyal
May 23, 2007·Journal of Medical Virology·Jacques R KremerClaude P Muller

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