Detection of hepatitis C virus transmission using DNA mass spectrometry.

Ganova-Raeva LM, Dimitrova ZE, Campo DS, Yulin L, Ramachandran S, Xia GL, Honisch C, Cantor CR, Khudyakov YE.

Source

Molecular Epidemiology and Bioinformatics Laboratory, Division of Viral Hepatitis, Centers for Disease Control and Prevention, 1600 Clifton Rd. NE, MS A-33, Atlanta, Georgia 30329, USA.

Abstract

The molecular detection of transmission of rapidly mutating pathogens such as hepatitis C virus (HCV) is commonly achieved by assessing the genetic relatedness of strains among infected patients. We describe the development of a novel mass spectrometry (MS)-based approach to identification of HCV transmissions. MS was used to detect products of base-specific cleavage of RNA molecules obtained from HCV PCR fragments. The MS-peak profiles (MSPs) were found to reflect variation in the HCV genomic sequence and the intra-host composition of the HCV population. Serum specimens (n=60) originating from case-patients of 14 epidemiologically confirmed outbreaks and unrelated controls (n=25) were tested. Neighbor-joining trees constructed using MSP-based Hamming distances showed 100% accuracy, and linkage networks constructed using a threshold established from the Hamming distances between epidemiologically unrelated cases showed 100% sensitivity and 99.93% specificity in transmission detection. The MS approach is rapid, robust, reproducible and cost-effective, and applicable to investigating transmissions of other pathogens.