Recombinant expression of Taenia solium TS14 antigen and its utilization for immunodiagnosis of neurocysticercosis.
In order to evaluate the potential use of TS14 antigen in an enzyme-linked immunosorbent assay (ELISA) for immunodiagnosis of neurocysticercosis (NC), its open reading frame (ORF) was amplified by RT-PCR from mRNA isolated from Taenia solium cysticerci. The ORF was subcloned into the expression vector pET-28a, and was used to transform Escherichia coli BL21 (DE3) cells to produce TS14 antigen. The His-tagged expressed protein was purified on a nickel affinity column. Using the HISTS14 as antigen, ELISA was positive for 100% of cerebrospinal fluid (CSF) and 97% of serum samples from NC patients. No positive results were observed with sera and CSF samples from control groups. Cross-reactivity with sera from patients with schistosomiasis and Chagas' disease was not observed. Serum samples from patients with taeniasis were evaluated and 2 of 13 cases showed reactivity in this assay. Our data indicate the usefulness of HISTS14 in ELISA for an accurate and rapid assay for diagnosis of NC and seroepidemiological studies.
da Silva
Marcia Ramos Monteiro
MR
Laboratório de Biologia Molecular, Departamento de Genética e Evolução, Universidade Federal de São Carlos, São Carlos, CEP 13565-905, SP, Brazil. marciamg@usp.br
Maia
Antônio Augusto Mendes
AA
Espíndola
Noeli Maria
NM
Machado
Luís dos Ramos
Ldos R
Vaz
Adelaide José
AJ
Henrique-Silva
Flávio
F
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood* / Q000134:cerebrospinal fluid* / Q000276:immunology; D000947:Antigens, Helminth / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D015801:Helminth Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D008970:Molecular Weight; D020019:Neurocysticercosis / Q000097:blood / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods; D013045:Species Specificity; D041201:Taenia solium / Q000276:immunology*
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015801:Helminth Proteins; D011994:Recombinant Proteins