Single-tube Protocol for Culture-independent Spoligotyping of Based on MeltArray

Overview

Journal J Clin Microbiol

Specialty Microbiology

Date 2023 Dec 19

PMID 38112521

Authors

Zihan Xia

Biyi Su

Chunxia Tu

Siyang Sun

Yaoju Tan

Ye Xu

Qingge Li

Affiliations

Soon will be listed here.

Abstract

Spacer oligonucleotide typing (spoligotyping), the first-line genotyping assay for (MTB), plays a fundamental role in the investigation of its epidemiology and evolution. In this study, we established a single-tube spoligotyping assay using MeltArray, a highly multiplex polymerase chain reaction (PCR) approach that runs on a real-time PCR thermocycler. The MeltArray protocol included an internal positive control, , to indicate the abundance of MTB via the quantification cycle and 43 spacers to identify the spoligotype via melting curve analysis. The entire protocol was completed in a single step within 2.5 hours. The lowest detectable copy number for the tested strains was 20 copies/reaction and thus sufficient for analyzing both culture and sputum samples. We conclude that MeltArray-based spoligotyping could be used immediately in low- and middle-income countries with a high tuberculosis burden, given its easy access, improved throughput, and potential applicability to clinical samples.

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