危重脑炎脑膜炎患儿的临床诊断与病原学检测分析

Clinical diagnosis and etiology detection of critical meningitis/encephalitis patient

目的对脑炎脑膜炎病例进行临床诊断与实验室检测,在病原学上为临床治疗提供直接依据。方法对入院患儿进行血常规、尿常规、生化及血凝检查,采集患儿脑脊液、血液及咽拭子标本,进行脑膜炎奈瑟菌、B型流感嗜血杆菌、金黄色葡萄球菌、肺炎链球菌、猪链球菌、大肠杆菌等分离培养;ELISA法检测麻疹病毒、风疹病毒IgM抗体;设计特异性引物RT-PCR法检测肠道病毒、乙脑病毒、水痘-带状疱疹病毒、单纯疱疹病毒、腮腺炎病毒,对核酸检测阳性的病原体进行基因测序、Blast比对及应用Mega 5.0构建分子进化树;根据病原体的种类及基因型别,有针对性地选择药物进行临床救治。结果脑炎脑膜炎患儿脑脊液、血液及咽拭子标本细菌培养均为阴性;咽拭子标本腮腺炎病毒PCR核酸检测阳性,琼脂糖凝胶电泳产物大小约300 bp;阳性产物测序结果与腮腺炎病毒疫苗株比较,存在部分核苷酸变异,关键的抗原位点没有发生变化;所测基因序列经分子进化分析确定为腮腺炎病毒基因F型,与目前中国流行毒株基因型别一致,患儿经对症治疗已康复出院。结论细菌学和病毒学实验室筛查,对引起脑炎脑膜炎的常见病原体具有积极的作用,早期病毒核酸测序能确诊腮腺炎病毒感染类别及基因型别。

【Objective】 To verify the clinical diagnosis and etiology detection for critical meningitis/encephalitis patient and then provide scientific basis for clinical therapy.【Methods】 Routine test of biochemistry,blood and urine were carried out as the patients were admitted.The samples of cerebrospinal fluid,blood and throat swab were collected for neisseria,haemophilus influenza,staphylococcus aureus,streptococcus pneumonia,streptococcus suis and escherichia coli bacterial cultures.The IgM of measles and rubella were detected by ELISA and specific primers were designed to detect the virus of enterovirus,Japanese encephalitis virus,herpes zoster virus,herpesvirus hominis and mumps virus by PCR.The positive product was verified by gene sequencing,blast and molecular evolution analysis.Appropriate drug for clinical therapy then was provided according to the genetype of the pathogen.【Results】 The results of bacterial cultures were negative,PCR product of the throat swab was positive for mumps virus and the product was about 300bp,the sequencing had partial nucleotide variation to the vaccine strain of mumps virus and the type was confirmed to the genetype F by evolution analysis.【Conclusion】 Bacteriology and virus laboratory detection can provide scientific basis for the common pathogen caused Meningitis/Encephalitis clinically,early phase viral nucleic acid detection can confirm the genetpye of the mumps virus.