婴幼儿轮状病毒两种检测方法的比较(1)
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[摘要] 目的：比较轮状病毒(RV)金标免疫层析法(GICA)和电镜法(EM)，为实验室合理选择检测方法提供依据。 方法：采用GICA和EM法对120例婴幼儿腹泻标本和100例正常婴幼儿粪便标本进行检测，计算阳性率，并对其检测结果进行统计学分析与比较。结果：EM法的阳性率37.5%(45/120)高于(GICA)法的阳性率33.3%(40/120)，差异有统计学意义(Kappa=0.392，χ2=19.64，P<0.01)。两种方法对正常对照组进行检测，结果均为阴性。结论：GICA法和EM法检测婴幼儿粪便中轮状病毒，结果有明显差异。EM法阳性率高，而GICA法低，并且两种方法的假阳性率几乎不存在。
[关键词] 轮状病毒；金标免疫层析法；电镜法；方法比较
[中图分类号] R446.5[文献标识码]B [文章编号]1673-7210(2011)04(a)-091-02
The comparison of two methods of infant rotavirus detection
SUN Anmin, MA Chunyan, LI Xianglei
Department of Clinical Laboratory, the First People's Hospital of Kaifeng City, He'nan Province, Kaifeng 475000, China
[Abstract] Objective: To carry out the gold standard immunochromatography assay (GICA) and the electron microscopy (EM) to compare the infant rotavirus (RV), which provides basis for a reasonable detecting method for laboratory. Methods: By using the GICA and the EM methods we selected 120 cases of diarrhea in infants and 100 normal samples of stool specimens for testing, and calculated the positive rates. The results were statistically analyzed and compared. Results: The positive rate by EM method (45/120, 37.5%) was higher than that by GICA method (40/120, 33.3%). The differences were significant (Kappa=0.392, χ2=19.64, P<0.01). We also detected the normal control group by these two methods, the results were negative. Conclusion: By the GICA and the EM methods to detect the rotavirus in infants, the results are significantly different. The positive rate of EM is high and of the GICA is low. And the false positive rate of the two methods is almost non-existent.
[Key words] Rotavirus; Gold standard immunochromatography; Electron microscopy; Comparison
轮状病毒(RV)是婴幼儿急性胃肠炎(腹泻)尤其是秋冬季腹泻的主要病因[1]，多为A群。新生儿RV感染也较普遍，一般通过粪-口途径传播，且多为无症状感染或轻型腹泻症状，常见于6个月~2岁的婴幼儿。通常使用酶联免疫吸附试验(ELISA)、聚丙烯酰胺凝胶电泳(PAGE)、金标免疫层析法(GICA)和电镜(EM)法来检测病毒，我院主要采用后两种方法。本文通过大量的实验数据，从实验的特异性和敏感度上对两种方法进行比较，以探讨两种不同方法在检测轮状病毒中的价值。现报道如下：
1 材料与方法
1.1 材料
1.1.1 仪器与试剂采用轮状病毒快速诊断试剂盒(胶体金法)，购自北京万泰生物药业有限公司；120 kV，JEM-1230透射电子显微镜，日本电子株式会社(JEOL)生产。
1.1.2 标本来源本院2008~2009年临床门诊和住院送检的腹泻婴幼儿大便标本120例，男70例，女50例，年龄0~2岁。另设正常对照组100例，年龄0~2岁，男60例，女40例，大便标本正常。
1.2 方法
1.2.1 大便标本的收集在症状出现以后尽快收集粪便标本，用清洁的塑料瓶子装盛。所需标本量约为1 g。如果不立即检测可在2℃~8℃冰箱保存1~2 d。长时间保存需要于-20℃冷冻，使用前彻底解冻并恢复到室温再检测。
1.2.2 GICA法检测在试管中加入0.5 ml稀释液，加适量粪便标本(固体直径约5 mm，液体50 μl)到试管中，使浓度为5%~10%，充分混匀，用滴管取2~3滴加入样品孔，5~10 min判读结果(必须在15 min以内判读结果 ......
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