Simple multiplex PCR assays to detect common pathogens and associated genes encoding for acquired extended spectrum betalactamases (ESBL) or carbapenemases from surgical site specimens in Vietnam

Abstract

Surgical site infection (SSI) is common in Vietnamese post-operative patients. It contributes to increased morbidity, mortality, hospitalization time and health care expenditure. Bacterial culture is considered the gold standard procedure to identify SSI pathogens and antibiotic resistant properties; however, it can detect microbes that can readily grow and is time-consuming. We propose optimized multiplex PCR assays to diagnose the most relevant microbes and associated genes encoding for acquired extended spectrum betalactamases (ESBL) or carbapenemases from Vietnamese patients with SSI in a hospital setting in Hanoi.MethodsNinety-one patients (n = 91) were collected in order to identify microbial pathogens and associated genes encoding for acquired extended spectrum betalactamases (ESBL) or carbapenemases by both conventional bacterial culture and in-house multiplex PCR assays.Result and conclusionThe novel in-house multiplex PCR assays are comparable to the bacterial culture approach in screening for common pathogens causing SSI and for relevant genotypes conferring betalactam/carbapenem resistance for bacteria. This is the first report of Turkey-specific ESBL gene (PER-1) and two Oxacilinase families (Oxa23 and Oxa 58) in Vietnam.

A survey of extended-spectrum beta-lactamase-producing Enterobacteriaceae in urban wetlands in southwestern Nigeria as a step towards generating prevalence maps of antimicrobial resistance

“…The presence of class A (Ambler classification scheme) ESBL genes (bla CTX-M , bla GES , bla PER , bla SHV , bla TEM , bla VEB ), class B ampC beta-lactamases (bla CMY-1, bla CMY-2, bla ACC, bla ACT, bla DHA and bla FOX ), and class D oxacillinase genes (bla OXA-1, bla OXA-2, bla OXA-10, bla OXA-23, bla OXA-24, bla OXA-48, bla OXA-51, bla OXA-58 ) was screened by PCR as previously described [35,36]. PCR primers and annealing temperatures are provided in S2 Table. Klebsiella pneumoniae strain Mnasey (bla CTX-M-15 , bla SHV-11 , bla TEM-1B ), Acinetobacter baumannii AYE (bla VEB ), ) and Providencia rettgeri H1736 (bla OXA-10 ) were used as positive controls.…”

The occurrence of ESBL-producing Escherichia coli carrying aminoglycoside resistance genes in urinary tract infections in Saudi Arabia

“…Molecular genotyping has been used concurrently with phenotyping techniques to detect and confirm antimicrobial drug resistance data and to detect Gram-negative ESBL producing bacteria [10]. PCR, multiplex PCR and oligonucleotide microarray-based assays have been developed and used to monitor the emergence of ESBLs and many other drug-resistant genes from E. coli , K. pneumonia and A. baumannii [11–15]. Strain characterization by multilocus sequence typing (MLST) is the method of choice in many clinical and research laboratories due to its high discriminatory power [16, 17].…”

Significance of nucleic acid testing in diagnosis and treatment of post-neurosurgical meningitis caused by multidrug-resistant Acinetobacter baumannii: a case report

“…Thus, we were unable to initiate an optimized antibiotic regimen. However, by using our in-house molecular assays [12], we could detect genetic material of A. baumannii . Moreover, the associated extended spectrum beta-lactamase ( ESBL ) encoding gene ( VEB ) and carbapenemase-encoding gene NDM-1 , and the metallo-beta-lactamase IMP [13] were also identified, suggesting multidrug resistance of A. baumannii .…”

“…The CSF culture was still negative for any microorganism. Therefore, in-house molecular assays were indicated [12], which led to the identification of genetic fragments of A. baumannii . Furthermore, the beta-lactam carbapenem resistance genes Vietnamese extended-spectrum beta-lactamase ( VEB ), OXA-58, New Delhi metallo-beta-lactamase-1 ( NDM-1 ), and Imipenemase ( IMP ), respectively, were detected (Fig.…”