A PCR-restriction enzyme technique for determining dengue virus subgroups within serotypes.
The polymerase chain reaction (PCR) and restriction enzyme analysis were used to develop a rapid and simple procedure for identifying geographic subgroups of dengue virus within serotypes for epidemiologic investigations. The entire structural protein region of dengue viruses was amplified and the products were digested with the endonucleases AluI or DdeI. By comparing the restriction fragment length polymorphisms (RFLPs), we recognized dengue-2 and dengue-3 subgroups that corresponded to those previously determined by oligonucleotide fingerprinting or genomic sequencing. This procedure can be performed in 2 days without the use of radioisotopes, and results can be interpreted without computer analysis. For those analyses which require only subgroup affiliations, this is a useful tool for rapidly screening multiple virus isolates.
Vorndam
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V
Division of Vector-Borne Infectious Diseases, National Centers for Infectious Diseases, Center for Disease Control and Prevention, San Juan, Puerto Rico 00921-3200.
Kuno
G
G
Rosado
N
N
D000330:Aedes; D000818:Animals; D002460:Cell Line; D004262:DNA Restriction Enzymes; D003715:Dengue / Q000821:virology; D003716:Dengue Virus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012703:Serotyping; D014773:Virology / Q000379:methods
D004262:DNA Restriction Enzymes