Establishment And Application Of Brucellosis, Lyme Disease Colloidal Gold Diagnostic Method

Brucellosis, an emerging zoonoses, has been shown to be prevalent to various degrees in developping countries. Due to the special pathogenic mechanism of Brucella, the existing methods of diagnosis of brucellosis have many problems. However, serological methods play important role in the detection and diagnosis of brucellosis. In this study, a fast and convenient immunochromatographic strip diagnostic method were established based on the serological techniques. Lyme disease, caused by tick-borne Borrelia burgdorferi, is a zoonosis. Due to the presence of Borrelia burgdorferi multiple genotypes, the patients and infected animals showed different clinical symptoms. Therefore, the clinical diagnosis is prone to misdiagnosis, leads to an increasing trend in recent years in natural foci, and danger to public health. Immunofiltration assay used was established in this study.The main work was carried out in this study:(1) Extraction and purification of B. melitensis 16 M LPS, identify its purity and concentration. Coated with LPS antigen, the sound GICA test strip technology and manufacturing conditions were optimized. Compared with the RBPT and SAT, the strips were used to detecting different sera both from animals and human samples. The results show that LPS immunochromatographic strip optimal coating concentration of 2.0 mg/m L. 440 serum samples test results. The GICA test strip showed 95.45% consistence with SAT, it was significantly higher than the RBPT showed 79.55% consistence with SAT. The positive rates of the GICA test strip was 84.32%, the positive rates of the SAT was 86.59%, there is no difference.(2) Based on the recombinant protein BmpA of Lyme which was constucted by our laboratory, identified its purity and concentration and coated with recombinant protein BmpA antigen, the sound immune infiltration cartridge and manufacturing conditions were optimized. The immune infiltration cartridge were used to detecting sera from patients and sheep with suspect infected Lyme. The results show that recombinant protein Bmp A immune infiltration cartridge optimal coating concentration was 2.0 mg/mL. The 3 positive sera has been detected from 8 serum which has identified to Lyme disease. There were 2 positive sera detected from the 30 sheep serum through percolation card, and the result of the percolation card is consistent with WB testing.This method was established by LPS as coating antigen immunochromatographic strip to diagnose brucellosis that can be used for sifting ranch brucellosis. Through optimization immunofiltration assay, it has been more convenient and faster detection than previous and lay the foundation for Lyme disease diagnose technology.