小儿隐孢子虫病的实验诊断研究  

Laboratory study on diagnosis of cryptosporidiosis in children with diarrhea

作　　者：郑成中[1] 李为明[2] 崔凤文[2] 崔守信[2] 杨颍 

机构地区：[1]解放军第三○六医院儿科,北京100101 [2]第三军医大学新桥医院儿科

出　　处：《中华儿科杂志》2000年第6期369-372,共4页Chinese Journal of Pediatrics

摘　　要：目的　探讨不同检测方法对小儿腹泻粪便中隐孢子虫卵囊 (CSO)的诊断效果。方法对 6 49例急性腹泻患儿的粪便采用 7种方法检测CSO :(1)番红 美兰染色法 (SMB) ;(2 )改良抗酸染色法 (MAF) ;(3)蔗糖漂浮 (SSF)后SMB染色法 ;(4)PBS 乙醚 (PBS E)离心沉淀后MAF染色法 ;(5 )聚蔗糖 泛影酸钠 (FSD)纯化后扫描电镜法 (SEM) ;(6 )FSD或Percoll纯化后免疫组化法 (ABC) ;(7)Percoll纯化后间接免疫荧光法 (IFA)。对 6 49例患儿的新鲜粪便标本同时采用前 4种方法检测 ,而对患儿的经预处理低温存放的 710份标本 (已知阳性 86份 ,已知阴性 6 2 4份 )同时采用 7种方法检测。结果　共检出隐孢子虫腹泻病 2 6例 ,阳检率 4.0 %。SMB和MAF(SMB +MAF)方法的敏感性、特异性分别为80 .2 %和 94.1%、6 1.6 %和 98.4% (P <0 .0 1) ,它们的检测准确性分别为 92 .4%和 93.9% (P >0 .0 5 ) ,SSF +SMB、PBS E法 +MAF法及ABC法的敏感性、特异性、准确性均为 10 0 % ,与SMB、MAF法相比较均有非常显著性意义 (P <0 .0 1)。结论　粪便直接涂片SMB法阳检率高 ,假阳性也高 ,适宜筛查 ;MAF法与之相反 ,漏诊率高 ;ABC法与金标准IFA法检测效果与IFA法一样 ,但方法繁琐造价高 ;SSF或PBS E法浓集CSO后SMB、MAF法的检测效果与IFA法相似 ,且经济、?Objective To evaluate the diagostic effects, sensitivity, specificity and accuracy of 7 different methods in detecting cryptosporidium oocysts (CSO) in fecal specimens of children with diarrhea. Methods Seven microscopy-based CSO detection methods were applied to unconcentrated and concentrated fecal smears from 649 children with acute diarrhea. The seven methods were as follows: (1) A direct hot safranin-methylene blue (SMB) stain method, (2)a direct modified acid-fast (MAF) stain method, (3)a modified Sheather′s sucrose flotation (SSF) plus SMB stain method, (4) a PBS-ether concentration (PBS-E) plus MAF stain, (5) a combination of Ficoll-sodium diatrizoate (FSD) discontinuous density gradient and observation under scanning electron microscope (SEM), (6) FSD or Percoll purifying CSO method plus an avidin-biotin-peroxidase complex (ABC) technique, and (7) a monoclonal antibody-based indirect immunofluorescence assay (IFA). The first 4 methods listed above were simultaneously used to examine CSO in fresh stool samples from 649 children with diarrhea. All seven methods were used in 710 previously collected, pretreated, stored (at 2-8℃) specimens (86 known positives, 624 known negatives) from 649 children with diarrhea. Results A total of 649 diarrhea cases were examined for fecal CSO, which were identified in 26 cases (4.0%). The sensitivity, specificity and accuracy of the SMB stain and the MAF (or SMB+MAF) method for the fecal CSO were 80.2% and 94.1%, 61.6% and 98.4% ( P < 0.01), 92.4% and 93.9% ( P >0.05), respectively. The sensitivity, specificity and accuracy of the SSF+SMB, PBS-E+MAF, ABC methods were all 100%, which were statistically significant ( P <0.01 for all) as compared to those of SMB, MAF or SMB+MAF methods. Conclusions The direct SMB staining of the unconcentrated fecal smears showed a higher positive rate than the one of the direct MAF stain, but its mis-diagnosis rate was higher as well. This test may be suitable for screening. The MAF stain had a high false-negative rate. T

关 键 词：隐孢子虫病 儿童 荧光免疫测定 单克隆抗体 

分 类 号：R725.3[医药卫生—儿科]