Establishmen And Comparative Of LAMP And Fluorescent Quantitative PCR For Brucella Detection

To detect and identify brucella in a rapid, accurate and convenient manner, we established and evaluated two measures--Fluorescence quantitative PCR and LAMP technology using different primers according to the principles of each method.Based on the conservative gene BCSP31of brucella, Fluorescence quantitative polymerase chain reaction (PCR) probe and primers are designed. LAMP uses a unique method to design primers through brucella inserting sequence IS711, which is a muLti-copy gene. The reaction system is optimized with the standard of plasmid and the lowest detection of two methods was tested. In order to evaluate the specificity of two methods, the bacteria and its similar species were tested through cross reaction and were amplified. To ensure the positive rate of specimen, we tested patient’s blood and serum specimen using fluorescence quantitative PCR and compared their positive rates. Serum was firstly used for brucella molecuLar biology detection nationwide. The resuLts showed that serum was more suitable for brucella molecuLar biological detection than whole blood. By comparison, both Fluorescent quantitative PCR and LAMP had high specificity and sensitivity. The lowest detection was within102copy/uL magnitude. In identifying50brucella strains from different places, data showed that the Fluorescence quantitative PCR, the conventional PCR and LAMP are in100%accordance with the traditional methods of brucella identification. Whereas in the test of clinical serum samples, the positive detection rates of these three methods were89%,62%and93%. The coincidence rate in the negative control was100%.Two methods are highly applicable in clinical diagnosis for brucellosis. However, the Fluorescence quantitative PCR has a higher requirement of special equipment and laboratory environment, thus more suitable for research institutes. The LAMP is more suitable for brucellosis epidemiological investigation and primary prevention given lower requirement of special equipment.