Detection and differentiation of human herpesviruses 1-5 by consensus primer PCR and RFLP.
Eight human viruses of the Herpesviridae family represent a significant public health problem world-wide. Detection and typing of five of the human herpesviruses (HSV-1, HSV-2, VZV, EBV, and CMV) was performed by applying a consensus primer polymerase chain reaction (PCR). The amplified PCR products from the five human herpesviruses were typed based on their restriction enzyme digestion polymorphism with Hinf I and Alu I. Fifteen clinically suspected specimens from herpesvirus-infected patients were also evaluated. A fragment of the DNA polymerase gene from each of the five human herpesviruses was successfully amplified by the set of consensus primers. Their amplicons obtained by PCR from the template DNAs were subjected to restriction endonuclease digestion and human herpesviruses 1-5 could be clearly differentiated and typed. This method can be used to detect and differentiate between the five human herpesviruses in clinical specimens. This study demonstrates the value of testing for five human herpesviruses by consensus PCR and restricted fragment length polymorphism (RFLP). These procedures are simple and straightforward techniques for the investigation of clinical specimens.
Sakulwira
Kamol
K
Inter-Department of Medical Microbiology, Faculty of Graduate Studies, Chulalongkorn University, Bangkok 10330, Thailand.
Vanapongtipagorn
Pijitra
P
Theamboonlers
Apiradee
A
Bhattarakosol
Parvapan
P
Wananukul
Siriwan
S
Poovorawan
Yong
Y
D001483:Base Sequence / Q000235:genetics; D002648:Child; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length
D004279:DNA, Viral