Universal Primer Design for the Detection of Diverged CTX-M Extended-Spectrum β-Lactamases (ESBL) That Give Penicillin and Cephalosporin Resistance During Superbug Infections
التفاصيل البيبلوغرافية
العنوان:
Universal Primer Design for the Detection of Diverged CTX-M Extended-Spectrum β-Lactamases (ESBL) That Give Penicillin and Cephalosporin Resistance During Superbug Infections
The complexity of diverged β-lactamases in multidrug-resistant bacteria has created a panic as common antibiotics failed to cure infections. We found >40%, >25%, and 0.002% of bacteria in Kolkata water bodies (river, sea, rain) were ampicillin, tetracycline, and meropenem resistant, respectively. More than 20 unique sequence classes of β-lactamases were known to cause MDR inactivating penicillin, cephalosporin, and carbapenem drugs. Further, there are other diverged mdr genes such as drug efflux genes (acrAB, mexAB-EF, tetA), drug-modifying genes (aacA1/C1, strA/B), and drug-binding genes (tetS/M, PBP, sul1) that were present in a single bacterial plasmid suggesting too many PCR reactions to be performed to understand the drug resistomes. Thus, we devised a method of reduction in PCR assays using seq-2 and forced multi-align analysis of >200 blaCTX-M protein sequences for universal primer design. The universal primer set recognized major type-1, type-2, and type-9 blaCTX-M mutants including clinically relevant blaCTX-M-15. We also made blaTEM-SHV common primers. This study was supported by the WHO recommendations to adopt a uniform policy for AMR detection and antibiotic therapy worldwide. Interestingly, we have also devised a solution (MDR-Cure) for the treatment of plasmid-borne MDR infections using Indian phytoextracts.
