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2024, Vol: 14, Issue: 1
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Research Article


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

Beauty Makamure, Tsitsi Bandason, Vinie Kouamou, Martha Chipinduro, Forget Makoga, Janice Martin, Isabel Mashita, Justin Mayini, Shungu Munyati, John Z Metcalfe, Jody Phelan, Pasipanodya Nziramasanga, Junior Mutsvangwa, Justen Manasa.

Abstract
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 carried out 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.

Key words: Deoxyribonucleic acid, Mycobacterium tuberculosis, storage temparature


 
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How to Cite this Article
Pubmed Style

Makamure B, Bandason T, Kouamou V, Chipinduro M, Makoga F, Martin J, Mashita I, Mayini J, Munyati S, Metcalfe JZ, Phelan J, Nziramasanga P, Mutsvangwa J, Manasa J. Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures. J Microbiol Infect Dis. 2023; 13(1): 31-37. doi:10.5799/jmid.1265384

Web Style

Makamure B, Bandason T, Kouamou V, Chipinduro M, Makoga F, Martin J, Mashita I, Mayini J, Munyati S, Metcalfe JZ, Phelan J, Nziramasanga P, Mutsvangwa J, Manasa J. Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures. https://www.jmidonline.org/?mno=302656984 [Access: February 03, 2024]. doi:10.5799/jmid.1265384

AMA (American Medical Association) Style

Makamure B, Bandason T, Kouamou V, Chipinduro M, Makoga F, Martin J, Mashita I, Mayini J, Munyati S, Metcalfe JZ, Phelan J, Nziramasanga P, Mutsvangwa J, Manasa J. Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures. J Microbiol Infect Dis. 2023; 13(1): 31-37. doi:10.5799/jmid.1265384


Vancouver/ICMJE Style

Makamure B, Bandason T, Kouamou V, Chipinduro M, Makoga F, Martin J, Mashita I, Mayini J, Munyati S, Metcalfe JZ, Phelan J, Nziramasanga P, Mutsvangwa J, Manasa J. Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures. J Microbiol Infect Dis. (2023), [cited February 03, 2024]; 13(1): 31-37. doi:10.5799/jmid.1265384


Harvard Style

Makamure, B., Bandason, . T., Kouamou, . V., Chipinduro, . M., Makoga, . F., Martin, . J., Mashita, . I., Mayini, . J., Munyati, . S., Metcalfe, . J. Z., Phelan, . J., Nziramasanga, . P., Mutsvangwa, . J. & Manasa, . J. (2023) Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures. J Microbiol Infect Dis, 13 (1), 31-37. doi:10.5799/jmid.1265384


Turabian Style

Makamure, Beauty, Tsitsi Bandason, Vinie Kouamou, Martha Chipinduro, Forget Makoga, Janice Martin, Isabel Mashita, Justin Mayini, Shungu Munyati, John Z Metcalfe, Jody Phelan, Pasipanodya Nziramasanga, Junior Mutsvangwa, and Justen Manasa. 2023. Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures. Journal of Microbiology and Infectious Diseases, 13 (1), 31-37. doi:10.5799/jmid.1265384


Chicago Style

Makamure, Beauty, Tsitsi Bandason, Vinie Kouamou, Martha Chipinduro, Forget Makoga, Janice Martin, Isabel Mashita, Justin Mayini, Shungu Munyati, John Z Metcalfe, Jody Phelan, Pasipanodya Nziramasanga, Junior Mutsvangwa, and Justen Manasa. "Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures." Journal of Microbiology and Infectious Diseases 13 (2023), 31-37. doi:10.5799/jmid.1265384


MLA (The Modern Language Association) Style

Makamure, Beauty, Tsitsi Bandason, Vinie Kouamou, Martha Chipinduro, Forget Makoga, Janice Martin, Isabel Mashita, Justin Mayini, Shungu Munyati, John Z Metcalfe, Jody Phelan, Pasipanodya Nziramasanga, Junior Mutsvangwa, and Justen Manasa. "Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures." Journal of Microbiology and Infectious Diseases 13.1 (2023), 31-37. Print. doi:10.5799/jmid.1265384


APA (American Psychological Association) Style

Makamure, B., Bandason, . T., Kouamou, . V., Chipinduro, . M., Makoga, . F., Martin, . J., Mashita, . I., Mayini, . J., Munyati, . S., Metcalfe, . J. Z., Phelan, . J., Nziramasanga, . P., Mutsvangwa, . J. & Manasa, . J. (2023) Assessment of Quantity and Quality of Multidrug-Resistant Tuberculosis DNA Extracts Stored at Different Temperatures. Journal of Microbiology and Infectious Diseases, 13 (1), 31-37. doi:10.5799/jmid.1265384


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