A molecular platform for the diagnosis of multidrug-resistant and pre-extensively drug-resistant tuberculosis based on single nucleotide polymorphism mutations present in Colombian isolates of Mycobacterium tuberculosis

Developing a fast, inexpensive, and specific test that reflects the mutations present in Mycobacterium tuberculosis isolates according to geographic region is the main challenge for drug-resistant tuberculosis (TB) control. The objective of this study was to develop a molecular platform to make a rapid diagnosis of multidrug-resistant (MDR) and extensively drug-resistant TB based on single nucleotide polymorphism (SNP) mutations present in the rpoB, katG, inhA, ahpC, and gyrA genes from Colombian M. tuberculosis isolates. The amplification and sequencing of each target gene was performed. Capture oligonucleotides, which were tested before being used with isolates to assess the performance, were designed for wild type and mutated codons, and the platform was standardised based on the reverse hybridisation principle. This method was tested on DNA samples extracted from clinical isolates from 160 Colombian patients who were previously phenotypically and genotypically characterised as having susceptible or MDR M. tuberculosis. For our method, the kappa index of the sequencing results was 0,966, 0,825, 0,766, 0,740, and 0,625 for rpoB, katG, inhA, ahpC, and gyrA, respectively. Sensitivity and specificity were ranked between 90-100% compared with those of phenotypic drug susceptibility testing. Our assay helps to pave the way for implementation locally and for specifically adapted methods that can simultaneously detect drug resistance mutations to first and second-line drugs within a few hours.

Physicochemical properties of water quality of Imeh, Edegelem and Chokocho communities located along Otamiri-oche River in Etche Ethnic Nationality of Rivers State, Nigeria

Some physiochemical properties of water quality of Otamiri-oche River which runs through Imeh, Edegelem and Chokocho Communities in Etche ethnic nationality of Rivers State, Nigeria were investigated. Samples were collected in triplicate from the three different locations along the river in both rainy and dry seasons. Each sampling points is located 500m away from the successive one, and the samples were taken from the open river at the zone extending beyond the shoreline. Samples were stored in ice packs and immediately taken to the laboratory to investigate some selected physico-chemical characteristics. Also, bacteriological study was carried out on the samples. The laboratory results of the selected physico-chemical parameters when compared with the world health organization (WHO) standard showed that all the selected physico-chemical parameters were within the world health organization (WHO) permissible in both rainy and dry season except pH. For rainy season, the pH values were 5.00±0.00, 5.40 ±0.40, and 5.40± 0.30 for Imeh, Edegelem and Chokocho respectively. For the dry season, the pH values were 5.83± 0.01, 5.56±0.01, 5.90±0.01 for Imeh, Edegelem and Chokocho respectively. Also magnesium hardness exceeded the World Health Organization (WHO) permissible limit in rainy season only. The results of bacteriological study showed that all the sampling locations contained high numbers of coliform bacteria in both rainy and dry seasons. The number of coliform bacteria during rainy season was 25.00±0.00, 25.00±0.00, and 35.00±0.00 for Imeh, Edegelem and Chokocho respectively. Those of dry season were 93.00±3.55, 56.66±4.49, and 35.60±4.17 for Imeh, Edegelem and Chokocho respectively. The difference between the rainy and dry season was tested statistically using the t-test. The results of the statistical analyses showed a significant difference (p<0.05) between the rainy and dry seasons.