COMPARISON OF DIRECT ELECTRON-MICROSCOPY, IMMUNE ELECTRON-MICROSCOPY, AND ROTAVIRUS ENZYME-LINKED IMMUNOSORBENT-ASSAY FOR DETECTION OF GASTROENTERITIS VIRUSES IN CHILDREN
COMPARISON OF DIRECT ELECTRON-MICROSCOPY, IMMUNE ELECTRON-MICROSCOPY, AND ROTAVIRUS ENZYME-LINKED IMMUNOSORBENT-ASSAY FOR DETECTION OF GASTROENTERITIS VIRUSES IN CHILDREN
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DOI：
10.1128/jcm.13.5.976-981.1981
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发表时间：
1981-01-01
影响因子：
9.4
通讯作者：
CHANOCK, RM
中科院分区：
文献类型：
--
作者：
BRANDT, CD;KIM, HW;CHANOCK, RM
411例患有急性胃肠炎的婴幼儿的第一个粪便样本在水中约有10%的悬浮物，在2分钟的阴性染色后用电镜检查。该方法能够通过免疫电子显微镜（IEM）和轮状病毒酶联免疫吸附试验（ELISA）等综合技术最终检测出199例轮状病毒感染病例中的88%、所有22例腺病毒感染病例和47%的15-27 nm病毒感染病例。204例粪便直接电磁检测感染中，76%在观察2 min内检测到，94%在观察6 min内检测到。1型和2型轮状病毒颗粒的可视化效率大致相同，尽管2型轮状病毒感染更为常见。在直接电镜检测病毒感染时，直肠拭子制剂明显不如粪便制剂。为了有效地显示直肠拭子标本中的病毒，需要进行IEM检查。ELISA法检测轮状病毒最灵敏；采用该方法可检出所有EM或IEM显示轮状病毒颗粒的感染病例，但在常规间接ELISA推定为轮状病毒阳性的1834份标本中，有73%的标本在EM、IEM、阻断型ELISA、确认型ELISA或这些方法联合的基础上出现假阳性。用改良的确认性ELISA检测粪便标本时，对同一动物的轮状病毒阴性（免疫前）抗体的稀释度低于轮状病毒阳性（免疫后）捕获抗体的稀释度，显然可以消除轮状病毒ELISA假阳性反应。
An approximate 10% suspension in water of the 1st available stool sample from 411 infants and young children with acute gastroenteritis was examined by EM after 2 min of negative staining. This procedure enabled the detection of 88% of the 199 rotavirus infections, all of the 22 adenovirus infections, and 47% of the 15-27 nm virus infections ultimately detected by a combination of techniques, including immune electron microscopy (IEM) and rotavirus enzyme-linked immunosorbent assay (ELISA). Of the 204 infections detected by direct EM of stools, 76% were detected within 2 min of viewing and 94% were detected within 6 min of viewing. Type 1 and type 2 rotavirus particles were visualized with approximately equal efficiency, although type 2 rotavirus infections were more common. Rectal swab preparations were clearly inferior to stool preparations for the detection of virus infection by direct EM. IEM examination was required for efficient visualization of viruses in rectal swab specimens. ELISA was the most sensitive method for the detection of rotaviruses; with this method, all infections in which rotavirus particles were visualized by EM or IEM were detected, but 73% of the 1834 specimens which were presumptively positive for rotavirus by conventional indirect ELISA were falsely positive on the basis of EM, IEM, blocking ELISA, confirmatory ELISA or a combination of these methods. False-positive rotavirus ELISA reactions apparently were eliminated when fecal specimens were tested in a modified confirmatory ELISA with a lower dilution of rotavirus-negative (pre-immunization) than rotavirus-positive (post-immunization) capture antibody from the same animal.