瓣膜赘生物组织均质处理后病原体培养方法研究  

Study on pathogen culture method after homogeneous pretreatment of heart vegetation tissue

作　　者：刘玉磊[1] 蒋晔 LIU Yulei;JIANG Ye(Department of Clinical Laboratory Center,Beijing Anzhen Hospital,Capital Medical University,Beijing 100029,China;Department of Clinical Laboratory,Tongren Hospital,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学附属北京安贞医院检验科,北京100029 [2]首都医科大学附属北京同仁医院检验科,北京100069

出　　处：《国际检验医学杂志》2021年第24期2953-2957,2962,共6页International Journal of Laboratory Medicine

摘　　要：目的探索组织均质前处理赘生物标本适宜条件,验证组织均质处理临床标本的使用效果,提高赘生物病原体检出率。方法首先通过定量培养,选择条件验证菌株的菌液稀释度。在不同温度、转速和时长条件下验证金黄色葡萄球菌、大肠埃希菌、近平滑念珠菌、戈登链球菌及艾肯菌均质处理后的存活数。选择菌株存活数无差异的均质程序进行菌株和赘生物组织的同时处理,确定组织破碎程度较高和菌株存活数较多的最佳条件。最后进行临床赘生物标本均质处理,将均质后的标本进行增菌分离培养和染色镜检后统计阳性率,与传统剪碎、研磨方法进行比对。结果1麦氏浊度的菌悬液取10μL进行定量培养时1∶1000稀释是适宜的稀释倍数;在温度为4℃、-20℃及-80℃,转速和时长分别为6 m/s 1 min、4 m/s 1 min 30 s、4 m/s 1 min条件下,验证菌株的存活数与对照菌液差异无统计学意义(P>0.05);在-20℃放置30 min,6 m/s 1 min条件下,组织破碎效果和菌株存活状况最佳;30份感染性心内膜炎患者心脏瓣膜赘生物直接培养阳性率为20.0%,剪碎培养阳性率为16.7%,研磨培养阳性率为20.0%,均质直接培养阳性率为23.3%,均质增菌培养阳性率为36.7%,均质增菌培养联合镜检阳性率为76.7%。结论均质处理临床心脏瓣膜赘生物标本的最佳条件是6 m/s 1 min、-20℃;均质处理后培养阳性率明显提高;组织均质前处理后,培养联合镜检的方法能够进一步提高病原体检出率;在组织病原体分离培养中,均质处理的方法是目前最佳的前处理方法,推荐临床验证和使用。Objective To explore the suitable conditions for pretreatment of cardiac vegetations by homogeneous,verify the effectiveness of the use of tissue homogeneous in treating clinical samples,and improve the detection rate of pathogens on cardiac vegetations samples.Methods Select appropriate dilution ratio of the experimental bacteria strains by quantitative culture.Verify the survival rate of Staphylococcus aureus,E.coli,Candida parapsilosis,Streptococcus gordon and Eikenella corrodens after homogenization under different temperature,speed and duration conditions.Pretreat the bacterial strains and the biological tissue under the program with on different survival number of the bacterial strains,find the best conditions for tissue fragmentation and higher survival rate of the strains.The clinical samples were homogenized by homogeneous.Bacterial culture and microscopic examination were carried out after homogenization.Positive rate were compared with the traditional methods,take cutting and grinding for example.Results The dilution rate of 1∶1000 was appropriate for the preparation the bacteria working solution for quantitative culture from 10μL bacteria solution of 1 Mc.Under the speed of 6 m/s 1 min,4 m/s 1 min 30 s,4 m/s 1 min at temperatures of 4℃,-20℃and-80℃conditions,there was no significant difference of the survival rate between the verified strains and the control.We got the best tissue crushing effect and survival number under-20℃froze for 30 min 6 m/s 1 min conditions.A total of 30 clinical cardiac vegetation samples were involved in this study.the positive rate of normal culture was 20.0%,culture after cut with scissors was 16.7%,culture after grinding was 20.0%,direct culture after homogenization was 23.3%,culture after enriching of the homogenized samples was 36.7%,homogeneous allied with microscopic examination was 76.7%.Conclusion The best conditions for homogeneous instrument treatment of clinical heart valve biopsy is 6 m/s 1 min,-20℃.The positive rate of culture after homogeneous treat

关 键 词：感染性心内膜炎 赘生物 均质 前处理 细菌培养 

分 类 号：R446.5[医药卫生—诊断学]