Biosafety and Proteome Profiles of Different Heat Inactivation Methods for Mycobacterium tuberculosis

Wang, Chenghui; Putri, Denise Utami; Lee, Jau-Ching; Liao, Chien-Chang; Tsao, Sung-tzu; Hsiao, Ai-Lin; Wu, Jhao-Hui; Chen, Xiaowei; Lee, Chih Hsin; Tsai, I‐Lin

doi:10.1128/spectrum.00716-21

Cited by 3 publications

(3 citation statements)

References 42 publications

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“…However, heat treatment has varying inactivation efficiencies, with some groups reporting uniform killing (Krugman 1975, Castro, Gonzalez et al 2009, and others raising issues with kill consistencies (Billard-Pomares, Bleibtreu et al 2019). Differences in temperature, duration and container type of heat inactivation has led to no uniform consensus being reached (Wang, Putri et al 2021). Ethanol can be used as a chemical inactivation method, with Dunne et al demonstrated that a ≥15minute incubation time was sufficient to inactivate a loop full of culture (Dunne, Doing et al 2014).…”

Section: Discussionmentioning

confidence: 99%

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A mycobacterial DNA extraction protocol designed for resource limited settings generates high quality whole genome sequencing

Percy,

Memelis,

Edwards

et al. 2024

Preprint

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Mycobacteria are major global human pathogens and includeMycobacterium tuberculosis, the causative agent of tuberculosis, andM. abscessus, an emerging multidrug resistant pathogen.M. abscessusaffects people with structural lung disease and those who are immunocompromised, most commonly causing pulmonary disease but also disseminated infections in the central nervous system and skin. High quality whole genome sequencing is essential to research mycobacterial epidemiology, pathogenesis and antimicrobial resistance. However, current DNA extraction protocols are time consuming, use toxic chemicals, require cold chain storage for certain reagents and can often result in poor quality, degraded DNA that directly impacts whole genome sequencing outputs. This is a particular challenge in low-income settings.Here, we report a novel optimised DNA extraction workflow forM. tuberculosisandM. abscessusthat invariably generates high quality Illumina short read sequencing data. We evaluated input culture CFU and physical cell disruption times. DNA quantity was determined using a Qubit fluorometer system with DNA integrity assessed using the Agilent TapeStation platform.We showed that this protocol facilitated complete genome assemblies ofM. abscessusandM. tuberculosisreference strains. There is no requirement for cold chain transport or storage of reagents, solvent extractions, or boiling to heat inactivate cultures, and the method does not require surfactant chemicals such as cetyltrimethylammonium bromide (CTAB).

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Section: Discussionmentioning

confidence: 99%

“…inactivation has led to no uniform consensus being reached(Wang, Putri et al 2021). Ethanol can be used as a chemical inactivation method, with Dunne et al demonstrated that a ≥15minute incubation time was sufficient to inactivate a loop full of culture(Dunne, Doing et al 2014).…”

mentioning

confidence: 99%

A mycobacterial DNA extraction protocol designed for resource limited settings generates high quality whole genome sequencing

Percy,

Memelis,

Edwards

et al. 2024

Preprint

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“…A sample inactivation step was performed to reduce the infection risk associated with clinical specimens that may contain MTB (17). Specimen material directly in contact with inactivation reagent (IR) for at least 1 hour was liqui ed and considered to have a reduced infection risk (18).…”

Section: Sample Size and Testing Proceduresmentioning

confidence: 99%

Performance evaluation of Abbott real-time PCR in the diagnosis of Mycobacterium tuberculosis in Addis Ababa, Ethiopia: A cross-sectional descriptive study

Tesema,

Berta,

Bitew

et al. 2023

Preprint

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Background In 2018, an estimated 10 million people developed tuberculosis, of whom more than 1.45 million died. The microscopy method used in most tuberculosis high burden and resource-limited countries is less accurate for diagnosing the disease. Thus, evaluation of the available diagnostic modalities in the country is crucial, and this study aimed to evaluate the performance of Abbott real-time PCR as a diagnostic technique for tuberculosis in Ethiopia. Methods A cross-sectional survey was conducted using sputum specimens collected from 150 presumptive tuberculosis patients from both public and private health facilities in Addis Ababa, Ethiopia, from May to June 2019. The laboratory investigation was conducted at the National Reference Laboratories of the Ethiopian Public Health Institute (EPHI). Results This finding indicated that 84.7% (127/150) and 61.3% (92/150) were smear and culture-negative, respectively. The overall diagnostic sensitivity of the Abbott real-time polymerase chain reaction (PCR) technique for the diagnosis of tuberculosis was 89.7% (52/58), that for smear-negative was 80.6% (29/36), and that for specificity was 92.4% (85/92). Drug resistance testing demonstrated diagnostic specificities of 87.5% and 100% for isoniazid and rifampicin, respectively, and a sensitivity of 92.3% for both. Conclusions This study demonstrated an outstanding performance of the Abbott real-time PCR technique for diagnosing tuberculosis using sputum specimens using culture as a reference standard. Thus, we recommend that Ethiopia's ministry and tuberculosis program implementers consider the Abbott real-time PCR technique for diagnosing tuberculosis and drug resistance testing, which is likely to be included in the national guidelines.

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Methods for the Inactivation of Mycobacterium tuberculosis: a Systematic Review of the Literature

Vigo,

Puyén,

Santos-Lázaro

et al. 2024

The International Journal of Mycobacteriology

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To systematize published laboratory methods to inactivate Mycobacterium tuberculosis (MTB) and to describe their effectiveness. We carried out a review of the scientific literature to identify the publications that reported methods for the inactivation of MTB, according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses recommendations. The search addressed inactivation methodologies used in Public health laboratories for the treatment of biological material and only included studies reporting the efficacy of the method. The database used were PubMed (National Library of Medicine) and LILACS (Latin American and Caribbean Literature in Health Sciences). We evaluated the quality of the studies with the JBI (Joanna Briggs Institute) Critical Instrument Appraisal Checklist. We included 14 publications meeting the established inclusion and exclusion criteria. These 14 studies actually described a total of 35 inactivation methods. Most of them combined heat treatment with some chemical or enzymatic agent, and there were very few applying a single strategy. Twenty-four (68.57%) methods demonstrated significant efficacy in inactivating MTB. The systematic review identified 35 methods of inactivation of MTB, published in 14 studies. Most of the methods combined physical treatment techniques, especially heat, with chemical and/or enzymatic treatment techniques, obtaining mostly good results in preventing the reproduction of the microorganism. PROSPERO (International Prospective Register of Ongoing Systematic Reviews) (Code CRD42024503621).

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Biosafety and Proteome Profiles of Different Heat Inactivation Methods for Mycobacterium tuberculosis

Cited by 3 publications

References 42 publications

A mycobacterial DNA extraction protocol designed for resource limited settings generates high quality whole genome sequencing

A mycobacterial DNA extraction protocol designed for resource limited settings generates high quality whole genome sequencing

Performance evaluation of Abbott real-time PCR in the diagnosis of Mycobacterium tuberculosis in Addis Ababa, Ethiopia: A cross-sectional descriptive study

Methods for the Inactivation of Mycobacterium tuberculosis: a Systematic Review of the Literature

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