FC-TRIPLEX Chagas/Leish IgG1: A Multiplexed Flow Cytometry Method for Differential Serological Diagnosis of Chagas Disease and Leishmaniasis

被引：13

作者：

Teixeira-Carvalho, Andrea ^{[1
]}

Freire Campos, Fernanda Magalhaes ^{[1
]}

Geiger, Stefan Michael ^{[2
]}

Rodrigues Rocha, Roberta Dias ^{[1
]}

de Araujo, Fernanda Fortes ^{[1
]}

Vitelli-Avelar, Danielle Marquete ^{[1
]}

Andrade, Marileia Chaves ^{[3
]}

Silva Araujo, Marcio Sobreira ^{[1
]}

Lemos, Elenice Moreira ^{[4
]}

de Freitas Carneiro Proietti, Anna Barbara ^{[5
]}

Sabino, Ester Cerdeira ^{[6
]}

Caldas, Rafaella Gaiotti ^{[1
]}

Camargos Freitas, Carolina Renata ^{[1
]}

Campi-Azevedo, Ana Carolina ^{[1
]}

Eloi-Santos, Silvana Maria ^{[7
,8
]}

Martins-Filho, Olindo Assis ^{[1
]}

机构：

[1] Fundacao Oswaldo Cruz, Ctr Pesquisas Rene Rachou, Lab Biomarcadores Diagnost & Monitoracao, Belo Horizonte, MG, Brazil

[2] Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Parasitol, Lab Helmintoses Intestinais, Belo Horizonte, MG, Brazil

[3] Univ Estadual Montes Claros, Ctr Ciencias Biol & Saude, Dept Fisiopatol, Montes Claros, Brazil

[4] Univ Fed Espirito Santo, Ctr Biomed, Nucleo Doencas Infecciosas, Vitoria, Brazil

[5] Fundacao Ctr Hematol & Hemoterapia Minas Gerais H, Serv Pesquisa, Belo Horizonte, MG, Brazil

[6] Univ Sao Paulo, Inst Med Trop, Dept Doencas Infecciosas, Sao Paulo, Brazil

[7] Univ Fed Minas Gerais, Fac Med, Dept Posgrad Patol Geral, Belo Horizonte, MG, Brazil

[8] Univ Fed Minas Gerais, Fac Med, Dept Propedeut Complementar, Belo Horizonte, MG, Brazil

来源：

PLOS ONE | 2015年 / 10卷 / 04期

关键词：

AMERICAN VISCERAL LEISHMANIASIS; POSTTHERAPEUTIC CURE ASSESSMENT; CRUZI IMMUNOGLOBULIN-G; TRYPANOSOMA-CRUZI; BLOOD-DONORS; CHEMOTHERAPEUTIC TREATMENT; SOUTHERN BRAZIL; ANTIBODIES; DEFINITION; PROGNOSIS;

D O I：

10.1371/journal.pone.0122938

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Differential serological diagnosis of Chagas disease and leishmaniasis is difficult owing to cross-reactivity resulting from the fact that the parasites that cause these pathologies share antigenic epitopes. Even with optimized serological assays that use parasite-specific recombinant antigens, inconclusive test results continue to be a problem. Therefore, new serological tests with high sensitivity and specificity are needed. In the present work, we developed and evaluated the performance of a new flow cytometric serological method, referred to as FC-TRIPLEX Chagas/Leish IgG1, for the all-in-one classification of inconclusive tests. The method uses antigens for the detection of visceral leishmaniasis, localized cutaneous leishmaniasis, and Chagas disease and is based on an inverted detuned algorithm for analysis of anti-Trypanosomatidae IgG1 reactivity. First, parasites were label with fluorescein isothiocyanate or Alexa Fluor 647 at various concentrations. Then serum samples were serially diluted, the dilutions were incubated with suspensions of mixed labeled parasites, and flow cytometric measurements were performed to determine percentages of positive fluorescent parasites. Using the new method, we obtained correct results for 76 of 80 analyzed serum samples (95% overall performance), underscoring the outstanding performance of the method. Moreover, we found that the fluorescently labeled parasite suspensions were stable during storage at room temperature, 4 degrees C, and -20 degrees C for 1 year. In addition, two different lots of parasite suspensions showed equivalent antigen recognition; that is, the two lots showed equivalent categorical segregation of anti-Trypanosomatidae IgG1 reactivity at selected serum dilutions. In conclusion, we have developed a sensitive and selective method for differential diagnosis of Chagas disease, visceral leishmaniasis, and localized cutaneous leishmaniasis.

引用

页数：15

共 29 条

[1] Evaluation of anti-lived and anti-fixed Leishmania (Viannia) braziliensis promastigote IgG antibodies detected by flow cytometry for diagnosis and post-therapeutic cure assessment in localized cutaneous leishmaniasis [J].

Alves Pereira, Valeria Rego ;

Reis, Luiza de Campos ;

Souza, Marina de Assis ;

de Oliveira, Andresa Pereira ;

Felinto de Brito, Maria Edileuza ;

Lage, Patricia S. ;

Andrade, Marileia Chaves ;

Rodrigues Rocha, Roberta Dias ;

Martins-Filho, Olindo Assis .

DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE, 2012, 74 (03) :292-298

[2] Anti-Trypanosoma cruzi Antibody Detection in Blood Donors in the Southern Brazil [J].

Araujo, A. B. ;

Vianna, E. E. S. ;

Berne, M. E. A. .

BRAZILIAN JOURNAL OF INFECTIOUS DISEASES, 2008, 12 (06) :480-482

[3] Conventional serological performance in diagnosis of Chagas' disease in southern Brazil [J].

Araujo, Anelise Bergmann ;

Aires Berne, Maria Elisabeth .

BRAZILIAN JOURNAL OF INFECTIOUS DISEASES, 2013, 17 (02) :174-178

[4] The use of IgG antibodies in conventional and non-conventional immunodiagnostic tests for early prognosis after treatment of Chagas disease [J].

Barbosa Wendling, Ana Paula ;

Vitelli-Avelar, Danielle Marquete ;

Sathler-Avelar, Renato ;

Geiger, Stefan Michael ;

Teixeira-Carvalho, Andrea ;

Gontijo, Eliane Dias ;

Eloi-Santos, Silvana Maria ;

Martins-Filho, Olindo Assis .

JOURNAL OF IMMUNOLOGICAL METHODS, 2011, 370 (1-2) :24-34

[5]

BRACHO CG, 1998, REV PANAM SALUD PUBL, V4, P94

[6] High Throughput Selection of Effective Serodiagnostics for Trypanosoma cruzi infection [J].

Cooley, Gretchen ;

Etheridge, R. Drew ;

Boehlke, Courtney ;

Bundy, Becky ;

Weatherly, D. Brent ;

Minning, Todd ;

Haney, Matthew ;

Postan, Miriam ;

Laucella, Susana ;

Tarleton, Rick L. .

PLOS NEGLECTED TROPICAL DISEASES, 2008, 2 (10)

[7] Anti-Trypanosoma cruzi immunoglobulin G1 can be a useful tool for diagnosis and prognosis of human Chagas' disease [J].

Cordeiro, FD ;

Martins-Filho, OA ;

Rocha, MOD ;

Adad, SJ ;

Corrêa-Oliveira, R ;

Romanha, AJ .

CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, 2001, 8 (01) :112-118

[8] Epidemiology, control and surveillance of Chagas disease-100 years after its discovery [J].

Coura, Jose Rodrigues ;

Pinto Dias, Joao Carlos .

MEMORIAS DO INSTITUTO OSWALDO CRUZ, 2009, 104 :31-40

[9]

da Saude MS-Ministerio, 2005, REV SOC BRAS MED S3, V38, P12

[10] The impact of Chagas disease control in Latin America - A review [J].

Dias, JCP ;

Silveira, AC ;

Schofield, CJ .

MEMORIAS DO INSTITUTO OSWALDO CRUZ, 2002, 97 (05) :603-612

← 1 2 3 →