Protein suspension chip method capable of quantitatively determining yersinia pestis and method for making same

Abstract

The invention aims at providing a protein suspension chip which can quantitatively detect the bacteria and a preparation method thereof, particulary relates to a protein suspension chip which is suitable for quantitative detection and analysis of Yersinia pestis (Y.pestis). The method has high sensitivity, strong specificity, good detection capacity and wide dynamic range, and establishes a new detection mode platform.

Description

technical field [0001] The invention relates to a protein suspension chip capable of quantitatively detecting bacteria and a preparation method thereof, in particular to a protein suspension chip suitable for quantitatively detecting and analyzing Yersinia pestis. Background technique [0002] Yersinia pestis (Yersinia pestis) is a Gram-negative, non-motile, non-spore-forming coccus bacillus, with Gimusa, Wright's or Wesson's staining showing bipolar dense staining, and can be infected at 4-40°C. For internal growth, the optimum growth temperature is 28-30°C, the optimum growth pH value is 7.2-7.6, and the upper and lower limits of pH tolerance are pH5.0 and pH9.6 respectively. Yersinia pestis evolved from pseudotuberculosis bacteria and is in an active period of evolution. Its main host is rodents, and it is mainly transmitted among animals through the bite of fleas. Occasionally, human diseases can be caused by flea bites or contact with animals and animal fur with Yersin...

AI Technical Summary

Problems solved by technology

The indirect hemagglutination method (IHA) for detecting Yersinia pestis specific antibody (F1 antibody) has low sensitivity and poor specificity (Williams, J.E., Arntzen, L., Robinson, D.M.&et al.Comparison of passive haemagglutination and enzyme-linkedimmunosorbent assay for serodiagnosis of plague. Bull WHO 1982, 60, 777-781.)

For the nucleic acid detection method, although PCR technology is used to detect and analyze Yersinia pestis (Hinnebusch, J.&Schwan, T.G. New method for plaguesurveillance using polymerase chain reaction to detect Yersiniapestis in fleas. J Clin Microbiol 1993, 31 (6), 1511 -1514.; Higgins, J.A., Ezzell, J., Hinnebusch, B.J., Shipley, M., Henchal, E.A. & Ibrahim, M.S. 5'nuclease PCR assay to detect Yersinia pestis. JClin Microbiol 1998, 36(8), 2284-2288 .; Neubauer, H., Meyer, H., Prior, J., Aleksic, S., Hensel, A. & Splettstosser, W. A combination of different polymerase chain reaction (PCR) assays for the presumptive identification of Yersinia pestis. J Vet MedB Infect Dis Vet Public Health 2000, 47(8), 573-580.), especially real-time fluorescent quantitative PCR technology (Herber t, T., Emil, C.R., Sascha, A.D.&etal.Rapid detection of Yersinia pestis with multiplex real-timePCR assays using fluorescent hybridisation probes. FEMS ImmunolMed Microbiol 2003, 38, 117-126), high specificity, not easy to be contaminated, but the methods of existing PCR technology are cumbersome, especially such methods include DNA / RNA extraction and complex operation steps, and cannot be used for the detection of non-nucleic acid substances such as proteins, so nucleic acid detection methods are limited and inconvenient

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