Development of a rapid and sensitive latex agglutination-based method for detection of group A rotavirus.
Considering the background of morbidity and mortality caused by human rotavirus, detection methods that use rotavirus group antigen (VP6) in either enzyme immunoassay (EIA) or latex agglutination test (LAT) has been employed routinely in clinical diagnostic and epidemiological studies. In order to develop a rapid and sensitive rotavirus group A LAT, part of segment 6 corresponding to conserved N-terminal portion of the VP6 (1-245 aa) was cloned in Escherichia coli expression pGEX vector (glutathione S-transferase-GST gene fusion system) that has been modified previously containing a histidine tail at C-terminus. The immunological propriety of the recombinant VP6 having a total molecular weight of 52 kDa was evaluated by Western blot and by the ability of inducing anti-recombinant VP6 polyclonal antibodies in rabbit. The polyclonal serum produced was conjugated to a latex support to detect rotavirus in stool specimens. The percentage values for sensitivity and specificity of the rotavirus group A LAT were 98.5% and 100%, respectively.
de Góes
Ana Carolina Magalhães Andrade
AC
Biopharmaceuticals Program, Bio-Manguinhos, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.
de Moraes
Marcia Terezinha Baroni
MT
de Castro Silveira
Waldemir
W
Araújo
Irene Trigueiros
IT
de H'alluin
Jean-Claude
JC
da Silva Souza
Waldemar
W
da Silva Junior
José Godinho
JG
Leite
José Paulo Gagliardi
JP
D000818:Animals; D000914:Antibodies, Viral / Q000302:isolation & purification; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology; D002648:Child; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005243:Feces / Q000821:virology; D015870:Gene Expression; D006801:Humans; D007841:Latex Fixation Tests / Q000379:methods*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012401:Rotavirus / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; C061332:VP6 protein, Rotavirus