Long‐Term Storage at −80°C: Effect on Rate of Recovery of <i>Mycobacterium tuberculosis</i> From Direct Acid‐Fast Bacilli Smear‐Positive Sputum Samples

Shinu, Pottathil; Singh, Varsha; Nair, Anroop B.; Farooq, Rumana; Ishaq, Sheikh

doi:10.1002/jcla.21904

Cited by 6 publications

(3 citation statements)

References 17 publications

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“…In this study, we found a high recovery rate of 80.2% from Mycobacteria Growth Indicator Tube (MGIT) cultures of tuberculosis isolates stored at ambient, 37 0 C, and -80 0 C temperatures. The recovery rate found in our study compared with a previous study, which reported an optimal recovery rate range of 63% to 90% after culturing neat sputum samples that had been stored at −80 0 C for three months) [22]. This is evident that isolates stored at not-so-ideal temperatures can still be recovered at rates comparable to those stored at ideal temperatures.…”

Section: Discussionsupporting

confidence: 62%

Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures

Makamure

2023

J Microbil Infect Dis

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Objectives: Globally, Multidrug-Resistant Tuberculosis (MDR-TB) remains a public health concern. Rapid identification of MDR-TB using conventional or novel technologies is crucial for effective treatment. Here we assessed the quantity and quality of DNA extracted from MDR-TB isolates, allowing whole-genome sequencing (WGS). Methods: This was a retrospective study on MDR-TB isolates from five studies conducted in Zimbabwe between 2011 and 2019. The isolates were stored under ambient 37 oC and -80 oC temperatures, respectively. These isolates were resuscitated and confirmed to be Mycobacterium tuberculosis (MTB). DNA was extracted using the N-Cetyl-N-trimethylammonium-bromide standard protocol. The concentration (A260nm) and purity (A260/280) of the extracted DNA before WGS (concentration ≥20ng/ul) were compared among the different storage conditions on Stata v15. Results: A total of 85 samples were successfully recovered from 106 retrieved. The overall recovery rate was 80.2%. We found a significant difference (p=0.005) in the concentration of the DNA samples by storage temperatures, with samples stored at -80 oC having the lowest concentration. Similarly, a significant difference (p=0.018) was found in the purity of the DNA (samples within the optimal range of 1.8 ± 0.2) by storage conditions, with 34/39 (87.2%) stored under ambient temperature, 18/20 (90.0%) stored in a 37 oC incubator and 1/4 (25.0%) stored at -80 oC. Conclusions: The better concentration and purity obtained from samples stored at 37 oC and ambient temperatures provide an impetus that such storage conditions could be used in many resource-limited settings where power supplies are a limitation to long storage conditions. J Microbiol Infect Dis 2023; 12(4):31-37.

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Section: Discussionsupporting

confidence: 62%

Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures

Makamure

2023

J Microbil Infect Dis

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“…Even at temperatures below freezing, the mycobacterial population freezes, but it survives. This is useful for long-term storage of mycobacteria for laboratory research [33,[137][138][139].…”

Section: Psychrophilic Mycobacterial Speciesmentioning

confidence: 99%

Nontuberculous Mycobacteria as Sapronoses: A Review

et al. 2022

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Mycobacteria are a unique group of microorganisms. They are characterised by exceptional adaptability and durability. They are capable of colonisation and survival even in very unfavourable conditions. In addition to the well-known obligate human pathogens, Mycobacterium tuberculosis and M. leprae, more than 200 other species have been described. Most of them form a natural part of the microflora of the external environment and thrive in aquatic and soil environments especially. For many of the mycobacterial species associated with human disease, their natural source has not yet been identified. From an ecological point of view, mycobacteria are saprophytes, and their application in human and animal diseases is opportunistic. Most cases of human disease from saprophytic mycobacteria occur in immunocompromised individuals. This adaptability and resilience to environmental pressures makes treatment of mycobacterial diseases (most often sapronoses and less often zoonoses) and permanent eradication of mycobacteria from the environment very difficult. Saprophytic mycobacterial diseases (sapronoses) are chronic and recurrent due to the fact of repeated endogenous or exogenous re-exposure. Therefore, knowledge regarding their occurrence in soil and dust would aid in the prevention of saprophytic mycobacterioses. In conjunction, their presence and ecological significance in the environment can be revealed.

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“…In the present study, LED FM microscopy could demonstrate a sensitivity and specificity of 76.8% and 99.19%, respectively. This higher sensitivity and specificity of LED FM direct microscopy can be implicated to various factors like patient inclusion criteria, burden of TB, quality of sputum and number of organisms present in sputum [22][23][24][25]. In fact, patients having cough for more than two weeks or longer periods were included in this study.…”

Section: Plos Onementioning

confidence: 99%

Papain-cetylpyridinium chloride and pepsin-cetylpyridinium chloride; two novel, highly sensitive, concentration, digestion and decontamination techniques for culturing mycobacteria from clinically suspected pulmonary tuberculosis cases

Shinu

Singh²,

Nair

et al. 2020

PLoS ONE

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Mycobacterial culture remains the gold standard for the diagnosis of tuberculosis. However, an appropriate digestion and decontamination method (DDM) is essential for the effective recovery of tubercle bacilli in culture. Therefore, the current study was designed to compare the performance of papain-cetylpyridinium chloride [papain-CPC] and pepsin-cetylpyridinium chloride [pepsin-CPC] DDMs against N-acetyl L-Cysteine-sodium hydroxide (NALC-NaOH) DDM for recovery of mycobacteria from clinically suspected pulmonary tuberculosis cases. To evaluate papain-CPC, pepsin-CPC and NALC-NaOH DDMs, sputum samples (N = 1381) were cultured on Lö wenstein-Jensen medium and the results were compared. The papain-CPC DDM showed sensitivity, specificity, positive predictive value, and negative predictive value of 100%, 93.27%, 71.7%, and 100%, respectively as compared to NALC-NaOH DDM. Similarly, pepsin-CPC DDM demonstrated sensitivity, specificity, positive predictive value and negative predictive value of 98.94%, 94.7%, 76.11%, and 99.81%, respectively. In summary, both papain-CPC and pepsin-CPC DDMs are highly sensitive and specific techniques for recovery of mycobacteria as compared to NALC-NaOH DDM. However, when the overall performances of all DDMs compared, papain-CPC DDM isolated increased number of mycobacterial isolates with comparatively higher numbers of colonies on LJ media than both pepsin-CPC and NALC-NaOH DDMs, indicating its potential to replace the NALC-NaOH DDM for recovery of mycobacteria from sputum samples.

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Long‐Term Storage at −80°C: Effect on Rate of Recovery of Mycobacterium tuberculosis From Direct Acid‐Fast Bacilli Smear‐Positive Sputum Samples

Cited by 6 publications

References 17 publications

Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures

Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures

Nontuberculous Mycobacteria as Sapronoses: A Review

Papain-cetylpyridinium chloride and pepsin-cetylpyridinium chloride; two novel, highly sensitive, concentration, digestion and decontamination techniques for culturing mycobacteria from clinically suspected pulmonary tuberculosis cases

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