Pilot study of a rapid and minimally instrumented sputum sample preparation method for molecular diagnosis of tuberculosis

Scientific Reports
Tanya M FergusonAngelika Niemz

Abstract

Nucleic acid amplification testing (NAAT) enables rapid and sensitive diagnosis of tuberculosis (TB), which facilitates treatment and mitigates transmission. Nucleic acid extraction from sputum constitutes the greatest technical challenge in TB NAAT for near-patient settings. This report presents preliminary data for a semi-automated sample processing method, wherein sputum is disinfected and liquefied, followed by PureLyse(®) mechanical lysis and solid-phase nucleic acid extraction in a miniaturized, battery-operated bead blender. Sputum liquefaction and disinfection enabled a >10(4) fold reduction in viable load of cultured Mycobacterium tuberculosis (M.tb) spiked into human sputum, which mitigates biohazard concerns. Sample preparation via the PureLyse(®) method and a clinically validated manual method enabled positive PCR-based detection for sputum spiked with 10(4) and 10(5) colony forming units (cfu)/mL M.tb. At 10(3) cfu/mL sputum, four of six and two of six samples amplified using the comparator and PureLyse(®) method, respectively. For clinical specimens from TB cases and controls, the two methods provided 100% concordant results for samples with 1 mL input volume (N = 41). The semi-automated PureLyse(®) method therefo...Continue Reading

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Citations

Nov 26, 2016·Transplant Infectious Disease : an Official Journal of the Transplantation Society·Sunil KumarDeepesh B Kenwar
Nov 9, 2018·Scientific Reports·Hyun-Ju ParkMin-Young Lee
Oct 3, 2019·Biosensors·Faye M Walker, Kuangwen Hsieh
Feb 11, 2020·ACS Applied Materials & Interfaces·Stephanie I PearlmanFrederick R Haselton
May 16, 2018·Analytical Chemistry·Andrew T BenderJonathan D Posner

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Methods Mentioned

BETA
PCR
nucleic acid amplification
nucleic acid extraction

Software Mentioned

OmniLyse
GeneXpert
PureLyse

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