PCR法对鸡卵黄中Coxiella burnetii菌的测定

Determination of Coxiella burnetii in egg yolk by PCR method

摘要:

【目的】通过用定量PCR加巢式PCR方法, 提高了对Coxiella burnetii (C. b) Com1基因的检出率; 通过对鸡卵中病原微生物Coxiella burnetii的基因检测, 明确鸡卵的食品安全性; 并对明确Coxiella burnetii的流行病学有重要意义。【方法】提取鸡卵DNA, 用定量PCR加巢式PCR方法检测上述基因, 并对PCR产物进行测序分析, 通过间接免疫荧光法观察鸡血白细胞中的微生物。【结果】用定量PCR加巢式PCR方法可检出4个以上的Coxiella burnetii Com1基因, 用此方法可测出鸡卵中Coxiella burnetii Com1基因达104?106个, 阳性率为5%?22%; 对阳性鸡卵Com1基因PCR产物的测序结果显示有变异菌株的存在; 免疫荧光法可见鸡卵中含有该微生物。【结论】由此认为鸡卵中存在病原微生物Coxiella burnetii, 可能是Q热传染源。

Abstract:

[Objective] Improve the detection rate of Coxiella burnetii (C. b) Com1 gene by quantitative PCR and nested PCR; determine the food safety of egg yolk by Coxiella burnetii Com1 gene detection, which has great magnificence for Coxiella burnetii epidemiology. [Methods] Extract DNA in chicken eggs, determine the Com1 gene by quantitative PCR and nested PCR, and sequence the PCR products. Observe the microorganism in chicken blood leukocytes by indirect immunofluorescence. [Results] Results showed that over 4 Com1 genes were detected by quantitative PCR and nested PCR, and the amount of Com1 genes in chicken eggs reached 104?106 with the positive rate of 5%?22%. The sequencing results of PCR products showed that there were variant strains in positive chicken eggs. Immunofluorescence observed the microorganism Coxiella burnetii in chicken eggs. [Conclusion] Therefore we draw the conclusion that the microorganism Coxiella burnetii exists in chicken eggs and it may be the source of infection of Q fever.