Publication

Sensitive and rapid detection of ganciclovir resistance by PCR based MALDI-TOF analysis

Journal Paper/Review - May 24, 2012

Units
PubMed
Doi

Citation
Zürcher S, Schaller A, Gorgievski-Hrisoho M, Garzoni C, Mohacsi P, Barbani M, Mühlemann K, Lüthi A, Mooser C, Flatz L. Sensitive and rapid detection of ganciclovir resistance by PCR based MALDI-TOF analysis. J Clin Virol 2012; 54:359-63.
Type
Journal Paper/Review (English)
Journal
J Clin Virol 2012; 54
Publication Date
May 24, 2012
Issn Electronic
1873-5967
Pages
359-63
Brief description/objective

BACKGROUND
Cytomegalovirus (CMV) infection is associated with significant morbidity and mortality in transplant recipients. Resistance against ganciclovir is increasingly observed. According to current guidelines, direct drug resistance testing is not always performed due to high costs and work effort, even when resistance is suspected.

OBJECTIVES
To develop a more sensitive, easy applicable and cost-effective assay as proof of concept for direct drug resistance testing in CMV surveillance of post-transplant patients.

STUDY DESIGN
Five consecutive plasma samples from a heart transplant patient with a primary CMV infection were analyzed by quantitative real-time polymerase chain reaction (rtPCR) as a surrogate marker for therapy failure, and by direct drug resistance detection assays such as Sanger sequencing and the novel primer extension (PEX) reaction matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) based method.

RESULTS
This report demonstrates that PEX reaction followed by MALDI-TOF analysis detects the A594V mutation, encoding ganciclovir resistance, ten days earlier compared to Sanger sequencing and more than 30 days prior to an increase in viral load.

CONCLUSION
The greatly increased sensitivity and rapid turnaround-time combined with easy handling and moderate costs indicate that this procedure could make a major contribution to improve transplantation outcomes.