Rapid detection of Epstein-Barr virus DNA by loop-mediated isothermal amplification method.
BACKGROUND
The loop-mediated isothermal amplification (LAMP) method is a novel technique for the amplification of specific DNA sequences.


OBJECTIVE
To establish the LAMP method for amplifying Epstein-Barr virus (EBV) DNA and to examine its reliability for the detection of EBV DNA in clinical specimens.


METHODS
Sera from 108 patients, who were initially suspected of primary EBV infection, were tested by the EBV LAMP method, and the results were compared with those of the real-time PCR assay. Serological examination was regarded as the standard diagnostic method.


RESULTS
To diagnose primary EBV infection, the sensitivity of LAMP was 86.4% and the specificity was 100%. The sensitivity of the real-time PCR assay was 84.1% and the specificity was 98.4%. Longitudinal analysis showed that the detection rate of EBV DNA in serum by the LAMP method decreased with time in accordance with the decrease of the EBV load. EBV DNA could not be detected in serum 40 days after onset of symptoms.


CONCLUSIONS
These results indicate that the sensitivity and specificity of the LAMP method are comparable to those of real-time PCR and that detecting EBV DNA in serum by this method is potentially useful for diagnosing primary EBV infection.
Iwata
Seiko
S
Department of Pediatrics, Anjo Kosei Hospital, Anjo, Japan.
Shibata
Yukiko
Y
Kawada
Jun-ichi
J
Hara
Shinya
S
Nishiyama
Yukihiro
Y
Morishima
Tsuneo
T
Ihira
Masaru
M
Yoshikawa
Tetsushi
T
Asano
Yoshizo
Y
Kimura
Hiroshi
H
D002675:Child, Preschool; D004279:DNA, Viral / Q000032:analysis* / Q000097:blood; D005260:Female; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010614:Pharynx / Q000821:virology; D012680:Sensitivity and Specificity
D004279:DNA, Viral