Authors' Contribution AS, FA and TMA designed the experiments. AS, MN and HA performed the experiments. TMA, JR and RA analyzed the data. All authors contributed in Manuscript writing.

GeneXpert MTB/RIF has revolutionized the tuberculosis diagnosis by simultaneous detection of Mycobacterium tuberculosis and resistance to RIF (rifampicin), a surrogate marker for multidrug-resistant TB in less than two hours. The RIF-resistance pattern in Balochistan, Pakistan, is not documented. This study was aimed to detect RIF-resistant TB and mutations in RNA polymerase beta (rpoB) gene of M. tuberculosis within 81-bp RRDR in Quetta, Pakistan using GeneXpert® MTB/RIF assay. In total, 2300 clinical specimens were collected from suspected TB patients at Fatima Jinnah General and Chest Hospital Quetta, Pakistan between January and August 2017. These specimens were analyzed by GeneXpert® MTB/RIF assay. The data was statistically analyzed using SPSS software. Out of 2300 clinical specimens, M. tuberculosis was positive in in 899 (39.1%) cases by GeneXpert® MTB/RIF assay [positive respiratory cases 42.9% (871/2032) and non-respiratory 10.4% (28/268) with statistically significant difference (χ2= 104.5, p<0.001)]. Among 899 MTB positive cases, 46 (5.1%) were RIF-resistance caused by various rpoB gene mutations within 81-bp RRDR. Most of the RIF-resistant isolates were observed to harbor mutationsin Probe E 78.3% (n=36) whereas mutations in Probe A, B, D were observed 2.2% (n=1), 4.3% (n=2), and 6.5% (n=3), respectively. However, none of cases had RIF-resistance associated with Probe C. Out of 46 RRD cases, 21 (45.7 %) were males and 25 (54.3 %) were females. Additionally, Xpert® test showed higher detection rate than fluorescent microscopy (39.1% vs 31.2%, P<0.05) and detected MTB in 186 (11.8%) smear-negative specimens. Among 42 confirmed TB patients had MDR contact and eight patients were co-infected with HIV. In conclusion, 5.1% of the TB patients showed rifampicin resistance. The most frequent rpoB genetic mutations were observed in codons 531/533 (Probe E, 78.3%) whereas the least within the sequence 511 (Probe A, 2.2%).

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Helicobacter pylori is a pathogenic bacterium causing gastric problems such as, peptic ulcers and stomach cancer. H. pylori were isolated from the stomach biopsy specimens (n = 100) of gastric patients by performing polymerase chain reaction (PCR) against cagA (cytotoxin associated gene A) and ureC (Urease subunit alpha) genes. Furthermore, antibiogram studies of the H. pylori isolates were evaluated against the common antibiotics. The overall detection rate of H. pylori was 71% in biopsy specimens of gastric patients. The antimicrobial susceptibility test revealed the resistance rate of H. pylori isolates against metronidazole (50%), clarithromycin (28.33%), tetracycline (21.66%), amoxicillin (18.33%), and ciprofloxacin (11.66%). However, the H. pylori isolates were completely resistant to vancomycin, erythromycin and nalidixic acid antibiotics. Clove oil showed a remarkable antimicrobial effect against H. pylori whereas, mild inhibition (10 mm) was observed in case of curcumin extract. Due to increase incident of resistance and high prevalence of H. pylori in gastric patients, natural antimicrobial like clove oil can be explored as an alternative treatment.