E-ISSN 2146-9369 | ISSN 2146-3158
 

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


 
ARTICLE TOOLS
Abstract
PDF Fulltext
How to cite this articleHow to cite this article
Citation Tools
Related Records
 Articles by Beauty Makamure
Articles by Tsitsi Bandason
Articles by Vinie Kouamou
Articles by Martha Chipinduro
Articles by Forget Makoga
Articles by Janice Martin
Articles by Isabel Mashita
Articles by Justin Mayini
Articles by Shungu Munyati
Articles by John Z Metcalfe
Articles by Jody Phelan
Articles by Pasipanodya Nziramasanga
Articles by Junior Mutsvangwa
Articles by Justen Manasa
on Google
on Google Scholar


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: October 13, 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 October 13, 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