title: Development of a rapid and sensitive latex agglutination-based method for detection of group A rotavirus.
abstract: 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.
authors.affiliation: Biopharmaceuticals Program, Bio-Manguinhos, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.
authors.forename: Ana Carolina Magalhães Andrade
authors.initials: AC
authors.lastname: de Góes
authors.forename: Marcia Terezinha Baroni
authors.initials: MT
authors.lastname: de Moraes
authors.forename: Waldemir
authors.initials: W
authors.lastname: de Castro Silveira
authors.forename: Irene Trigueiros
authors.initials: IT
authors.lastname: Araújo
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: de H'alluin
authors.forename: Waldemar
authors.initials: W
authors.lastname: da Silva Souza
authors.forename: José Godinho
authors.initials: JG
authors.lastname: da Silva Junior
authors.forename: José Paulo Gagliardi
authors.initials: JP
authors.lastname: Leite
mesh_terms: 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
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; C061332:VP6 protein, Rotavirus