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Pilot study of a rapid and minimally instrumented sputum sample preparation method for molecular diagnosis of tuberculosis
- Source :
- Scientific Reports
- Publication Year :
- 2016
- Publisher :
- Nature Publishing Group, 2016.
-
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 >104 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 104 and 105 colony forming units (cfu)/mL M.tb. At 103 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 "Equation missing"1 mL input volume (N = 41). The semi-automated PureLyse® method therefore performed similarly to a validated manual comparator method, but is faster, minimally instrumented and can be integrated into TB molecular diagnostic platforms designed for near-patient low-resource settings.
- Subjects :
- 0301 basic medicine
medicine.medical_specialty
Tuberculosis
030106 microbiology
Real-Time Polymerase Chain Reaction
Sputum sample
Article
Mycobacterium tuberculosis
03 medical and health sciences
Tuberculosis diagnosis
medicine
Humans
Sample preparation
Tuberculosis, Pulmonary
Colony-forming unit
Multidisciplinary
Chromatography
biology
business.industry
Sputum
Reproducibility of Results
Nucleic acid amplification technique
biology.organism_classification
medicine.disease
Surgery
medicine.symptom
business
Nucleic Acid Amplification Techniques
Subjects
Details
- Language :
- English
- ISSN :
- 20452322
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....b25eecdb65796d9521bf26ff47eba929
- Full Text :
- https://doi.org/10.1038/srep19541