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Comparison of Solid-phase Immune Electron Microscopy by Use of Protein A with Direct Electron Microscopy and Enzyme-linked Immunosorbent Assay for Detection of Rotavirus in Stool

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Specialty Microbiology
Date 1983 Nov 1
PMID 6315772
Citations 20
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Abstract

A total of 525 stool specimens collected during 1 year were examined for the presence of rotavirus by direct electron microscopy (EM), enzyme-linked immunosorbent assay (ELISA), and a solid-phase immune electron microscope method (SPIEM) utilizing protein A-coated grids for anchoring of specific viral antisera. Rotavirus was seen in 187 specimens; SPIEM detected 183 (97.8%), whereas direct EM and ELISA detected 161 (86%) and 166 (88.7%), respectively. No false-positive reactions were seen by ELISA. The sensitivity of the methods was evaluated by coded investigation of a dilution series of a positive sample, with a negative fecal specimen as diluent. SPIEM was approximately 30 times more sensitive than direct EM and 10 times more sensitive than ELISA. A study was done to compare the elapsed time for recognition of rotavirus by SPIEM and EM in 25 randomly selected positive specimens. All virus-positive specimens were detected within 2 min by SPIEM, whereas up to 9 min was required for direct EM. SPIEM with protein A is a highly sensitive method, useful for rapid detection of viruses in clinical specimens. Due to the direct visualization of virus particles by electron microscopy, there is no requirement for monospecific antisera for the method.

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References
1.
Derrick K . Quantitative assay for plant viruses using serologically specific electron microscopy. Virology. 1973; 56(2):652-3. DOI: 10.1016/0042-6822(73)90068-8. View

2.
Svensson L, von Bonsdorff C . Solid-phase immune electron microscopy (SPIEM) by use of protein A and its application for characterization of selected adenovirus serotypes. J Med Virol. 1982; 10(4):243-53. DOI: 10.1002/jmv.1890100404. View

3.
Bishop R, Davidson G, HOLMES I, Ruck B . Detection of a new virus by electron microscopy of faecal extracts from children with acute gastroenteritis. Lancet. 1974; 1(7849):149-51. DOI: 10.1016/s0140-6736(74)92440-4. View

4.
Kapikian A, Kim H, Wyatt R, Rodriguez W, Ross S, Cline W . Reoviruslike agent in stools: association with infantile diarrhea and development of serologic tests. Science. 1974; 185(4156):1049-53. DOI: 10.1126/science.185.4156.1049. View

5.
Middleton P, Petric M, Hewitt C, Szymanski M, Tam J . Counter-immunoelectro-osmophoresis for the detection of infantile gastroenteritis virus (orbi-group) antigen and antibody. J Clin Pathol. 1976; 29(3):191-7. PMC: 476020. DOI: 10.1136/jcp.29.3.191. View