Heat denaturation increases the sensitivity of the cytomegalovirus loop-mediated isothermal amplification method

Ryota Suzuki; Masaru Ihira; Yoshihiko Enomoto; Hiroaki Yano; Fumio Maruyama; Nobuhiko Emi; Yoshizo Asano; Tetsushi Yoshikawa

doi:10.1111/j.1348-0421.2010.00236.x

Heat denaturation increases the sensitivity of the cytomegalovirus loop-mediated isothermal amplification method

Microbiol Immunol. 2010 Aug;54(8):466-70. doi: 10.1111/j.1348-0421.2010.00236.x.

Authors

Ryota Suzuki¹, Masaru Ihira, Yoshihiko Enomoto, Hiroaki Yano, Fumio Maruyama, Nobuhiko Emi, Yoshizo Asano, Tetsushi Yoshikawa

Affiliation

¹ Department of Pharmacy, Faculty of Clinical Engineering, Fujita Health University School of Health Sciences, Japan.

PMID: 20646211
DOI: 10.1111/j.1348-0421.2010.00236.x

Abstract

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.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Cytomegalovirus / genetics
Cytomegalovirus / isolation & purification*
Cytomegalovirus Infections / virology*
DNA, Viral / genetics
Hot Temperature
Humans
Nucleic Acid Amplification Techniques / methods*
Sensitivity and Specificity

Substances

DNA, Viral