PCR法对鸡卵黄中Coxiella burnetii菌的测定

目的]通过用定量PCR加巢式PCR方法,提高了对Coxiella burnetii (C.b)CoMl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 Coml基因,用此方法可测出鸡卵中Coxiella burnetii Coml基因达104-106个,阳性率为5％-22％;对阳性鸡卵Coml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.

Determination of Coxiella burnetii in egg yolk by PCR method

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.