![]() There are also novel methods for the detection of Klebsiella pneumoniae, which are demonstrated to be more accurate and sensitive, like automatic bacteria identification system and mass spectrometry. ![]() PCR technique variants, such as multiplex PCR and real-time PCR methods, are primary diagnostic tools in hospitals and laboratories. Previous studies have established polymerase chain reaction (PCR)-based assays for human and fur-bearing animal (mink, raccoon dog, fox) Klebsiella pneumoniae. By contrast, genotype-based techniques are rapid and highly sensitive. Conventional phenotype-based laboratory techniques for the diagnosis of Klebsiella pneumoniae include pure culture, microscopic determination, biochemical examination are labor-intensive, time-consuming and easily interferred by other bacteria, even though regarded as the gold standards. īacterial detection generally depends on phenotype or genotype. Thus, an efficient, sensitive, and robust Klebsiella pneumoniae antibody detection method is highly desired to guide the prevention, intervention and control of the spread of Klebsiella pneumoniae in goat-farming industry. Seriously, Klebsiella pneumoniae-caused diseases and other secondary pathogenic bacteria infections eventually result in the death of goats. However, in this process, overuse and even abuse of antibiotics contribute to an increase in the number of outbreaks of Klebsiella pneumoniae. In China, along with increasing demand of goat livestock, goat husbandry industry has gradually adopted high-density breeding and fast fattening using concentrated feeding pattern. Klebsiella pneumoniae is an opportunistic pathogen parasitizing on the respiratory or intestinal tract of human and animals, and probably causes zoonotic disease such as meningitis, pneumonia, urinary tract inflammation, and even sepsis in the clinical veterinary, contributing to enormous potential threats to both human health and livestock production. Taken together, we successfully established an indirect ELISA method for detecting antibodies against Klebsiella pneumoniae in goats, which can be applied in production. In the detection of clinical veterinary samples, the positive rate of indirect ELISA was 6.74%, higher than that of conventional agglutination assays. ![]() The average coefficients of variation of intra-assay and inter-assay were 4.37 and 5.17% indicating favorable reproducibility of indirect ELISA. Furthermore, there was no cross-reaction between the coated antigen and goat serum positive for antibodies against other bacteria, indicating that indirect ELISA could detect Klebsiella pneumoniae specifically in most cases. The cut-off value was determined to be 0.28, and the analytical sensitivity was 1:800 (dilution of a positive sample). The optimal dilution buffers of bacterial antigen, the antibodies, and the blocking buffer were 0.05 mol/L carbonate buffer, 1% BSA phosphate buffer, and 1.5% BSA carbonate buffer, respectively. The optimal condition of coating and blocking were both 4 ☌ for 12 h. After screening and optimizing the conditions for detection, we determined the optimal working dilutions of the coated-bacterial antigen, the polyclonal antibody, and the enzyme-labeled secondary antibody that were 1:800 (2.99 × 10 7 CFU/ml), 1:6400, and 1:5000, respectively. In this study, we developed a Klebsiella pneumoniae goat polyclonal antibody and established an indirect ELISA method to detect the Klebsiella pneumoniae. However, there are little systematic methods for detection of goat Klebsiella pneumoniae in livestock production. Notably, infected goats may develop pneumonia, septicemia, which can lead to occasional death, resulting in great economic losses in goat-farming industry. Klebsiella pneumonia, a Gram-negative bacterium belonging to the genus Enterobacter, causes many human and livestock diseases.
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