产气荚膜梭菌qPCR法在动物模型和临床病例中的应用  

Detection of clostridium perfringens with qPCR in mouse models and a clinical case

作　　者：石玉玲[1] 徐少珊[1] 孙朝晖[1] 陈丽丹[1] 唐玲玲[1] 

机构地区：[1]广州军区广州总医院检验科,广州510010

出　　处：《中国医师杂志》2011年第3期324-327,共4页Journal of Chinese Physician

基　　金：基金项目：全军科学技术十一五计划专项基金资助项目（082009）;国家863项目生物高技术带来的生物安全危害应急技术

摘　　要：目的 利用产气荚膜梭菌qPCR法检测A型产气荚膜梭菌（Clostridium perfringens type A）感染小鼠的后肢的气性坏疽动物模型和一例阳性病例,以验证此法在早期诊断优势.方法 小鼠随机分成4组,10只/组分别肌肉注射3.5×109、3.5×108和3.5×107cfu/ml浓度的A型产气荚膜梭菌（ATCC13124）菌液0.1 ml,空白对照组肌肉注射生理盐水0.1 ml,建立小鼠感染的动物模型,利用产气荚膜梭菌qPCR方法 对动物模型及一例病例进行检测分析.结果 不同浓度菌液组（3.5×109、3.5×108、3.5×107cfu/ml实验组）和对照组在肌肉注射72 h内的死亡率为：90%、70%、10%和0;各组平均存活时间（hh：mm）依次为：20∶43±11∶12、37∶24±25∶39、68∶36±10∶45和72∶00±0∶00,各组荧光定量Ct值均值为：21.21±2.69、28.45±2.74、32.49±2.87和0.00±0.00,组间比较差异均有统计学意义（P＜0.05）.利用该法检测病者两处分泌物,Ct值为30.67、30.44.结论 利用产气荚膜梭菌qPCR法成功检测出未发病的小鼠中的产气荚膜梭菌,并成功的应用于一例病例中.Objective To detect of clostridium perfringens by qPCR in mouse models and a clinical case in order to offer early diagnosis.Methods 40 Kunming mice were randomly grouped and intramuscular injected clostridium perfringens type A in leg 0.1 ml（3.5 × 109cfu/ml or 3.5 × 108cfu/ml or 3.5× 107cfu/ml,diluted with saline）,while control group was injected with 9% sodium chloride 0.1ml.The mouse models and a clinical case were detected by qPCR.Results The death rate of 3.5 × 109,3.5 × 108,3.5 × 107cfu/ml and the blank group were 90%,70%,10% and 0% after intramuscular injection for 72 h spectively.The mean Ct values among these groups were 21.21 ±2.69,28.45 ±2.74,32.49 ±2.87 and 0.00 ± 0.00（P 〈 0.05）.The Ct values of the patient were 30.67 and 30.44.Conclusions Cclostridium perfringens could be successful identified with qPCR in mouse models when the mice still did not show any symptoms.

关 键 词：梭菌 产气荚膜/分离和提纯 聚合酶链反应/方法 气性坏疽/诊断 

分 类 号：R378.8[医药卫生—病原生物学]