In house reverse membrane hybridisation assay versus GenoType MTBDRplus and their performance to detect mutations in the genes rpoB, katG and inhA

Memórias do Instituto Oswaldo Cruz

Sergio Luiz Montego Ferreira Junior¹  Elis Regina Dalla Costa¹  Paula Gonçalves Dos Santos¹  Harrison Magdinier Gomes¹  Marcia Susana Nunes Silva¹  Leonardo Souza Esteves¹  Martha Maria Oliveira¹  Raquel De Abreu Maschmann¹  Afrânio Lineu Kritski¹  Philip Noel Suffys¹  Maria Lucia Rosa Rossetti¹ 

关键词: Mycobacterium tuberculosis;    resistance;    hybridisation;   

DOI  :  10.1590/0074-0276130469

来源: SciELO

【 摘 要 】

Drug-resistant tuberculosis (TB) threatens global TB control and is a major public health concern in several countries. We therefore developed a multiplex assay (LINE-TB/MDR) that is able to identify the most frequent mutations related to rifampicin (RMP) and isoniazid (INH) resistance. The assay is based on multiplex polymerase chain reaction, membrane hybridisation and colorimetric detection targeting of rpoB and katG genes, as well as the inhA promoter, which are all known to carry specific mutations associated with multidrug-resistant TB (MDR-TB). The assay was validated on a reference panel of 108 M. tuberculosis isolates that were characterised by the proportion method and by DNA sequencing of the targets. When comparing the performance of LINE-TB/MDR with DNA sequencing, the sensitivity, specificity and agreement were 100%, 100% and 100%, respectively, for RMP and 77.6%, 90.6% and 88.9%, respectively, for INH. Using drug sensibility testing as a reference standard, the performance of LINE-TB/MDR regarding sensitivity, specificity and agreement was 100%, 100% and 100% (95%), respectively, for RMP and 77%, 100% and 88.7% (82.2-95.1), respectively, for INH. LINE-TB/MDR was compared with GenoType MTBDRplus for 65 isolates, resulting in an agreement of 93.6% (86.7-97.5) for RIF and 87.4% (84.3-96.2) for INH. LINE-TB/MDR warrants further clinical validation and may be an affordable alternative for MDR-TB diagnosis.

【 授权许可】

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