TY - JOUR
T1 - Measles virus genotyping by nucleotide-specific multiplex PCR
AU - Kremer, Jacques R.
AU - Fack, Fred
AU - Olinger, Christophe M.
AU - Mulders, Mick N.
AU - Muller, Claude P.
PY - 2004/7
Y1 - 2004/7
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=3142691125&partnerID=8YFLogxK
UR - https://pubmed.ncbi.nlm.nih.gov/15243053
U2 - 10.1128/JCM.42.7.3017-3022.2004
DO - 10.1128/JCM.42.7.3017-3022.2004
M3 - Article
C2 - 15243053
AN - SCOPUS:3142691125
SN - 0095-1137
VL - 42
SP - 3017
EP - 3022
JO - Journal of Clinical Microbiology
JF - Journal of Clinical Microbiology
IS - 7
ER -