PCR法对鸡卵黄中Coxiella burnetii菌的测定

Determination of Coxiella burnetii in egg yolk by PCR method

【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。

[Objective] Improve the detection rate of Coxiella burnetii （C. b） Coral gene by quantitative PCR and nested PCR; determine the food safety of egg yolk by Coxiella burnetiiCoral gene detection, which has great magnificence for Coxiella burnetii epidemiology. IMethodsl Extract DNA in chicken eggs, determine the Coml 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 Coml genes were detected by quantitative PCR and nested PCR, and the amount of Coral genes in chicken eggs reached 104-10（＇ with the positive rate of 5%-22%. The sequencing results of PCR prod- ucts showed that there were variant strains in positive chicken eggs. Immunofluorescence ob- served the microorganism Coxiella burnetii in chicken eggs. [Conelusion] 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.