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

Authors : Beauty MAKAMURE

Pages : 31-37

Doi:10.5799/jmid.1265384

View : 9 | Download : 3

Publication Date : 2023-03-15

Article Type : Research Paper

Abstract :Objectives: Globally, Multidrug-Resistant Tuberculosis insert ignore into journalissuearticles values(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 insert ignore into journalissuearticles values(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 insert ignore into journalissuearticles values(MTB);. DNA was extracted using the N-Cetyl-N-trimethylammonium-bromide standard protocol. The concentration insert ignore into journalissuearticles values(A260nm); and purity insert ignore into journalissuearticles values(A260/280); of the extracted DNA before WGS insert ignore into journalissuearticles values(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 insert ignore into journalissuearticles values(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 insert ignore into journalissuearticles values(p=0.018); was found in the purity of the DNA insert ignore into journalissuearticles values(samples within the optimal range of 1.8 ± 0.2); by storage conditions, with 34/39 insert ignore into journalissuearticles values(87.2%); stored under ambient temperature, 18/20 insert ignore into journalissuearticles values(90.0%); stored in a 37 oC incubator and 1/4 insert ignore into journalissuearticles values(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; 12insert ignore into journalissuearticles values(4);:31-37.
Keywords : Deoxyribonucleic acid, Mycobacterium tuberculosis, storage temparature