摘要
目的 比较不同检测技术对不同时期采集的发热伴血小板减少综合征（SFTS）病例血清标本的检测结果,观察病毒核酸、IgM抗体、IgG抗体的动态特征,为疾病的诊断方法选择提供理论依据.方法 采集SFTS疑似病例早期和确诊病例康复后血清标本,采用实时荧光定量PCR技术和酶联免疫吸附实验（ELISA）分别检测病毒特异性核酸和IgM抗体、IgG抗体,分析比较各种方法的检出结果及与采集时间的关系,观察病毒核酸及特异性抗体的动态特征.结果 87份SFTS疑似患者血清,病毒核酸阳性率为53.41％,IgM抗体阳性率为31.03％,IgG抗体阳性率为3.41％.其中55份SFTS确诊病例标本,病毒核酸和IgM抗体检测结果的一致率为36.36％,两种方法检测结果差异有统计学差异（x2 =6.82,P=0.009）,kappa值=-0.257.核酸检测阳性标本采集与发病时间间隔均在12d以内,其中7～9d采集标本阳性检出率最高（100％）,经统计学分析,不同时间采集标本,核酸检测阳性率差异有统计学意义（x2=10.35,P=0.016）.34份SFTS恢复期血标本,核酸检测均为阴性,IgM抗体阳性率为41.18％,与急性期（38.23％）比较差异无统计学意义（P=1.00）.IgG抗体阳性率为94.12％,明显高于急性期IgG抗体阳性率（0％）.IgM和IgG抗体动态特征观察发现,IgM抗体在发病第2天即可检出,最晚检出时间为发病后第74天,30～60 d时间组吸光度均值和抗体检出率最高.IgG抗体最早检出时间为发病后第12天,最晚检出时间为发病后100 d.30 d内采集标本IgG抗体检出率仅为2.78％,30～ 60 d时间组IgG抗体检出率为100％.结论 发病2周内采集的SFTS疑似患者血标本,可优先考虑使用实时荧光PCR方法等检测病毒核酸或检测IgM抗体.IgM抗体虽可在发病2d后检出,但检出高峰出现时间多较晚,因此检测阴性并不能排除诊断.IgG抗体在恢复期标本中阳转率高,可作为疾病诊断的辅助手段.
Objective Compare the detection result of blood samples of severe fever with thrombocytopenia syndrome （SFTS） patients using different detection techniques,and observe the dynamic characteristics of the virus specific RNA,IgM antibody and IgG antibody,to provide theoretical basis for selection of diagnostic methods of disease.Methods Acute phase serum of suspected SFTS cases and convalescent serum samples of lab-confirmed cases were collected.Real-time fluorescence quantitative PCR and enzyme-linked immunosorbent assay （ELISA） were used to detect the virus specific RNA,IgM antibody and IgG antibody.The detection results of different methods,the relationship between positive results and the acquisition time,and the dynamic characteristics of viral nucleic acid and antibodies were analyzed.Results A total of 87 serum samples of the suspected SFTS patients were collected,the positive rate of virus specific RNA,IgM antibody and IgG antibody were 53.41%,31.03% and 3.41%,respectively.Among 55 confirmed cases of SFTS,the consistent rate of virus specific RNA and IgM antibody detection methods was 36.36%,and the difference between the two methods was significant （x2 =6.82,P =0.009）,kappa =-0.257.The sampling intervals of RNA positive samples were all within 12 days,of which the positive detection rate was highest after 7-9 days,and the difference was statistically significant （x2 =10.35,P =0.016）.In 34 SFTS convalescent serum samples,all the nucleic acid tests were negative,the positive rate of IgM antibody was 41.18%,which was not significantly different from the acute phase serum samples （P =1.00）.The positive rate of IgG antibody was 94.12%,which was significantly higher than that of acute IgG antibody （0%）.The dynamic characteristics of IgM and IgG antibody showed that IgM antibody could be detected on the second day after onset,the latest detection time was 74 days after onset,and the highest absorbance value and antibody detection rate occurred in 30-60 days.The earliest detection time of IgG antibody was 12 days after onset,and the last detection time was 100 days.The detection rate of IgG antibody and absorbance value increased rapidly after 30 days,and maintained in a high level.The detection rate of IgG antibody was 100% in 30-60 days.Conclusions Blood samples taken from SFTS suspected patients within two weeks of onset may be prioritized for detection of viral nucleic acids using Real-time fluorescence PCR or for detection of IgM antibodies by ELISA.Although IgM antibody can be detected 2 days after the onset,the peak appeared much later,so the negative result can＇t rule out the diagnosis.IgG antibody has a high seroconversion rate in convalescent samples,and can be used as an auxiliary tool for disease diagnosis.
作者
姜晓林
张晓梅
逢博
孙大鹏
王显军
丁淑军
Jiang Xiaolin, Zhang Xiaomei, Pang Bo, Sun Dapeng, Wang Xianjun, Ding Shujun(Department of Viral Disease Control and Prevention, Shandong Provincial Center for Disease Control and Prevention, Jinan 250014,Chin)
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
2018年第1期38-42,共5页
Chinese Journal of Experimental and Clinical Virology
基金
山东省自然科学基金（ZR2014HP030）
山东省科技发展计划（2012GHZ30031）
国家自然科学基金（81102171）
关键词
发热伴血小板减少综合征
SFTS病毒
核酸
抗体
荧光定量PCR
酶联免疫吸附实验
Severe fever with thrombocytopenia syndrome
SFTS virus
Nucleic acids
Antibodies
Real-time fluorescent quantitative PCR
Enzyme-linked immunosorbent assay
