Simple Assay for Detection of the Central Asia Outbreak Clade of the Mycobacterium tuberculosis Beijing Genotype

Abstract

The Central Asia outbreak (CAO) clade is a branch of the <i>Mycobacterium tuberculosis</i> Beijing genotype that is associated with multidrug resistance, increased transmissibility, and epidemic spread in parts of the former Soviet Union. Furthermore, migration flows bring these strains far beyond their areas of origin. We aimed to find a specific molecular marker of the Beijing CAO clade and develop a simple and affordable method for its detection. Based on the bioinformatics analysis of the large <i>M. tuberculosis</i> whole-genome sequencing (WGS) data set (<i>n</i> = 1,398), we identified an IS<i>6110</i> insertion in the <i>Rv1359-Rv1360</i> intergenic region as a specific molecular marker of the CAO clade. We further designed and optimized a multiplex PCR method to detect this insertion. The method was validated <i>in silico</i> with the recently published WGS data set from Central Asia (<i>n</i> = 277) and experimentally with <i>M. tuberculosis</i> isolates from European and Asian parts of Russia, the former Soviet Union, and East Asia (<i>n</i> = 319). The developed molecular assay may be recommended for rapid screening of retrospective collections and for prospective surveillance when comprehensive but expensive WGS is not available or practical. The assay may be especially useful in high multidrug-resistant tuberculosis (MDR-TB) burden countries of the former Soviet Union and in countries with respective immigrant communities.