Viability study of a multiplex diagnostic platform for Chagas disease.
A new multiplex assay platform was evaluated to detect Trypanosoma cruzi infection using the recombinant antigens CRA, FRA, CRAFRA fusion and parasite lysate. The antigens presented different sensitivity and specificity in a singleplex test when compared to a serial dilution of two pools comprising 10 positive serum samples and one pool of 10 negative samples. The recombinant protein CRA presented lower sensitivity (55%) in contrast to the 100% specificity and sensitivity of FRA, CRAFRA and T. cruzi lysate. These antigens also showed good results in a duplex test and the duplex test with CRAFRA/T. cruzi lysate showed better performance with 100% specificity and sensitivity, as well as a lower cut-off value in comparison to the other duplex test, FRA/T. cruzi lysate. Hence, when the antigens were used in duplex format, both tests showed decreased cut-off values and no interference between different bead sets, resulting in increasing sensitivity and specificity. The results of these multiplex tests show that they could be an alternative to singleplex detection for Chagas disease, and also indicate the necessity of using multiplex diagnostic tools to increase the sensitivity and specificity for diagnostic tests. Emerging data from the T. cruzi genome and from its ORFeome project will also allow the identification of new antigens for this disease detection application.
Foti
Leonardo
L
Instituto Carlos Chagas, Fiocruz, Curitiba, PR, Brasil.
Fonseca
Bruna de Paula Fonseca e
Bde P
Nascimento
Lilian Dias
LD
Marques
Christiane de Fatima Silva
Cde F
da Silva
Edmilson Domingos
ED
Duarte
Cesar Augusto Barros
CA
Probst
Christian M
CM
Goldenberg
Samuel
S
Pinto
Antônio Gomes
AG
Krieger
Marco Aurélio
MA
D000953:Antigens, Protozoan; D016022:Case-Control Studies; D014355:Chagas Disease / Q000175:diagnosis*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008863:Microspheres; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000953:Antigens, Protozoan; D011994:Recombinant Proteins