Direct identification by PCR of EBV types and variants in clinical samples.
Both Epstein-Barr virus (EBV) type A and type B, and variants of type A, were identified simultaneously by polymerase chain reaction (PCR) amplification of a DNA region coding for a 13 amino acid repeat in the Epstein-Barr virus nuclear antigen (EBNA) 6. Whereas this region varies extensively in type A isolates, no variation was seen in type B isolates. When a repetitive region in the LMP1-coding region was amplified by PCR, it was possible to distinguish individual variants of type B isolates from each other. Forty-two saliva samples from HIV-1-carrying individuals were examined for the presence of type A and type B virus. Both types and multiple variants of each type were found with a much higher frequency than in the saliva samples from healthy individuals. Type A EBV alone was detected in mouthwash samples from 6 infectious mononucleosis (IM) patients. Both type A and B were detected in the peripheral blood B-lymphocytes (PBL) from 1 healthy individual. The same type A variant was demonstrated both in PBL and in the mouthwash sample from another healthy individual. In this study it was shown that a combination of the EBNA 6- and LMP 1-specific PCRs followed by Southern hybridisation can be used to identify both type A and type B virus, as well as to distinguish between multiple variants of the same strain, in saliva and B-cells from both healthy and immunosuppressed individuals.
Falk
K I
KI
Microbiology and Tumorbiology Center, Karolinska Institute, Stockholm, Sweden.
Zou
J Z
JZ
Lucht
E
E
Linde
A
A
Ernberg
I
I
D002460:Cell Line; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics*; D014644:Genetic Variation; D015658:HIV Infections / Q000821:virology; D015497:HIV-1; D004854:Herpesvirus 4, Human / Q000145:classification / Q000235:genetics*; D006801:Humans; D007244:Infectious Mononucleosis / Q000097:blood / Q000821:virology*; D009067:Mouthwashes; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D012463:Saliva / Q000821:virology; D012680:Sensitivity and Specificity; D014763:Viral Matrix Proteins / Q000235:genetics*
C037662:EBV-associated membrane antigen, Epstein-Barr virus; D019309:Epstein-Barr Virus Nuclear Antigens; D009067:Mouthwashes; D014763:Viral Matrix Proteins