High throughput detection of drug-resistance gene mutations in HBV using MALDI-TOF mass spectrometry / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To develop a high-throughput clinical method on drug-resistance gene mutations of HBV using MALDI-TOF-MS.</p><p><b>METHOD</b>Using MassArray Assay Design software designed the iPLEX primers and followed the iPLEX instruction for amplification, SAP reaction, primer extenction, desalination, dispensing, MALDI-TOF-MS screening and data analysis of the gene mutation locus. 138 serum samples of chronic HBV patients with single drug-resistance or multiple drug-resistance on Lamivudin, adefovi, Entecavir were detected.</p><p><b>RESULT</b>The HBV gene mutation platform was successfully developed and applied on the high-throughput dectection of clinical serum samples. It was also a high throughput assay which could be used to detect for more than 138 samples once. The MALDI-TOF-MS technology and the DNA sequencing simultaneously examine 33 samples, in which result of 10 sample is inconsistent, the including 2 samples by MALDI-TOF-MS technology has not tested, 1 sample has 2 inconsistent mutations.</p><p><b>CONCLUSION</b>Detection of HBV gene mutations using MALDI-TOF-MS is highly-sensitive, highly-accurate, high-throughput, fast achieved and suitable to use in the diagnosis and monitoring of HBV.</p>