Heat denaturation increases the sensitivity of the cytomegalovirus loop-mediated isothermal amplification method.
A new protocol for CMV LAMP with an additional heat denaturation step was developed. While the sensitivity of the original CMV LAMP method was 500 copies/tube, sensitivity was increased by up to 100 copies/tube by additional heat denaturation. CMV DNA was detected in 103 of 350 samples (29.4%) by the original CMV LAMP procedure and 148 of 350 samples (42.3%) by the new CMV LAMP protocol. When the pp65 antigenemia assay was used as the standard method, the sensitivity, specificity, PPV, and NPV of the new protocol were 92.9%, 77.7%, 62.2%, and 96.5%, respectively.
Suzuki
Ryota
R
Department of Pharmacy, Faculty of Clinical Engineering, Fujita Health University School of Health Sciences, Japan.
Ihira
Masaru
M
Enomoto
Yoshihiko
Y
Yano
Hiroaki
H
Maruyama
Fumio
F
Emi
Nobuhiko
N
Asano
Yoshizo
Y
Yoshikawa
Tetsushi
T
D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000821:virology*; D004279:DNA, Viral / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity
D004279:DNA, Viral