THE RELEVANCE OF MICROBIOLOGICAL METHODS FOR THE DETECTION AND EARLY DIAGNOSIS OF CLIBSIELLA
Keywords:
Klebsiella pneumonia, digital PCRAbstract
The most frequent pathogen responsible for hospital respiratory tract infections and epidemics is Klebsiella pneumoniae (K. pneumoniae). Clinical diagnosis frequently uses the gold standard methods of microscopic inspection and biochemical identification, which have the drawbacks of low sensitivity, time commitment, and the need for complex equipment. To support clinical medicine and disease control, a sensitive, specific, and effective on-site identification of K. pneumoniae based on nucleic acid amplification is required. In order to specifically detect Klebsiella pneumoniae in fecal samples, this study set out to create a quick and sensitive droplet digital PCR (ddPCR) technique. Its clinical applicability was assessed by contrasting it with real-time PCR assays and traditional microbiological culture. A probe and particular primers were created to target the K. pneumoniae hemolysin (khe) gene. The specificity of the primers and probe was assessed using thirteen additional diseases. To evaluate the sensitivity, repeatability, and reproducibility of the ddPCR, a recombinant plasmid carrying the khe gene was created and examined using real-time PCR, ddPCR, and traditional microbial culture techniques. In comparison to real-time PCR, ddPCR demonstrated reduced inhibitory inhibition in the fecal sample. Thus, we developed a ddPCR-based assay technique for K. pneumoniae that is both sensitive and efficient. It might be a helpful technique for detecting K. pneumoniae in feces, and it might be a trustworthy way to find the bacteria that are causing the illness and assist direct treatment choices.This thorough investigation offers important insights into the region's K. pneumoniae epidemiological patterns and antibiotic susceptibility profile. The study's conclusions emphasize how urgently antimicrobial stewardship initiatives are needed to address the growing problem of antibiotic resistance. Healthcare providers can choose the right medications and enhance patient outcomes by having a better understanding of the K. pneumoniae resistance landscape. These statistics can help physicians make informed decisions about antibiotic treatment and aid in the creation of regional antibiotic policies.
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