Typing of human group A rotavirus with alkaline phosphatase-labeled oligonucleotide probes or a monoclonal enzyme immunoassay in unfrozen stools of children with diarrhea in Bangkok.
In developed countries, serotypes (or G types) have been identified in > 70% of group A rotavirus using monoclonal enzyme immunoassays (MEIAs); however, these assays have identified < 50% of rotavirus G types from developing countries presumably because the VP7 antigens were damaged by freezing and thawing during transportation of specimens. The VP7 (G) serotypes of rotavirus in unfrozen stool collected from children with acute diarrhea in Bangkok were determined using MEIA and compared to hybridization with alkaline phosphatase-labeled oligonucleotide probes. Reverse transcription of dsRNA coding for VP7 followed by polymerase chain reaction amplification of cDNA was used as an additional step prior to hybridization for 98 specimens that did not hybridize with the oligonucleotide probes. Of 251 rotavirus specimens, 208 (83%; 99% Cl = 76-89%) hybridized with G type specific oligonucleotides compared to 146 (58%; 99% Cl = 50-66%) that were typeable by MEIA. Forty-five (82%) of 55 stools containing G type 1, 80 of 84 (95%) containing G type 2, 0 of 3 containing G type 3, and 2 of 4 (50%) containing G type 4 as identified by MEIA hybridized with G type specific oligonucleotides. Differences in nucleotide sequences coding for VP7, in addition to destruction of the VP7 antigen by freezing and thawing of the specimen, may explain why not all rotavirus hybridized with G type specific probes.
Nirdnoy
W
W
Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand.
Sethabutr
O
O
Sakulkaipeara
T
T
Harikul
S
S
Hoge
C W
CW
Echeverria
P
P
D000469:Alkaline Phosphatase; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D002213:Capsid / Q000235:genetics; D036022:Capsid Proteins; D002675:Child, Preschool; D015342:DNA Probes; D003967:Diarrhea / Q000821:virology*; D005243:Feces; D006801:Humans; D007124:Immunoenzyme Techniques; D015336:Molecular Probe Techniques; D016133:Polymerase Chain Reaction / Q000379:methods; D012401:Rotavirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000821:virology*; D013785:Thailand
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D036022:Capsid Proteins; D015342:DNA Probes; C050779:VP7 protein, Rotavirus; D000469:Alkaline Phosphatase