Design multiplex PCR for detection of rapid and correct the metallobetalactamase

Abstract

One of the most important health and medical problems in many parts of the world is the spread of infectious diseases and the emergence of extent drug-resistant and innovation such as ESBL. Identification of them requires newer methods. One of the methods for identifying antibiotic resistance classes is Multiplex PCR. The purpose of this study is to identify ESBLs resistant gene using Multiplex PCR technique.In this study, three pair primers for ESBL (TEM, AmpC, KPC) were designed with bioinformatics software. Then, with control positive gene samples set-up of PCR was performed. Finally, additional experiments were performed with negative control samples and 50 isolates of the Escherichia coli isolated from Baqiyatallah hospital. The specificity and sensitivity of designed primers were also investigated.The results of the study showed that primers designed for MBL (SIM, GIM, IND) genes were able to simultaneously identify positive samples containing these genes. The sensitivity of Multiplex PCR for ESBL genes was 1 × 10–14. Also, the results showed that genotypic and phenotypic assays of clinical isolates of Escherichia coli were compatible with each other and the designed primers had good sensitivity (1 pg) and specificity (100%).This study showed that a Multiplex PCR designed with a sensitivity of 1 × 10–14 and a 100% specificity can correctly and fatly detect ESBL genes. Therefore, can suggest appropriate drug and antibiotic resistance to physicians determine the correct drug choice.