Molecular diagnosis of symptomatic toxoplasmosis: a 9-year retrospective and prospective study in a referral laboratory in São Paulo, Brazil

Author(s):Show less -

Camilo, Lilian Muniz ^[1] ; Pereira-Chioccola, Vera Lucia ^[1] ; Gava, Ricardo ^[1] ; Meira-Strejevitch, Cristina da Silva ^[1] ; Vidal, Jose Ernesto ^{[2, 3, 4]} ; Brandao de Mattos, Cinara Cassia ^[5] ; Frederico, Fabio Batista ^[6] ; De Mattos, Luiz Carlos ^[5] ; Junqueira Franco Spegiorin, Ligia Cosentino ^{[5, 7]} ; Antunes Murata, Fernando Henrique ^[8] ; Ferreira, Marina Neves ^{[9, 8]} ; Ulisses Barbosa, Deusenia Machado ^[8] ; Goncalves, Fausto da Silva ^[7] ; Dias, Cristiane Moraes ^[7] ; Catelan, Marcia Wakai ^[7] ; Siqueira, Rubens Camargo ^[8] ; Previato, Mariana ^{[8, 10]} ; Barbosa, Amanda Pires ^{[6, 11]} ; Cavallini, Danilo ^[6] ; Grp, FAMERP Toxoplasma Res

Affiliation:Show less -

^[1] Adolfo Lutz Inst, Ctr Parasitol & Micol, Sao Paulo, SP - Brazil

^[2] Univ Sao Paulo, Hosp Clin, Fac Med, Sao Paulo, SP - Brazil

^[3] Inst Infectol Emilio Ribas, Sao Paulo, SP - Brazil

^[4] Univ Sao Paulo, Inst Med Trop, Lab Invest Med LIM 4, Sao Paulo, SP - Brazil

^[5] Fac Med Sao Jose de Rio Preto, Sao Jose Do Rio Preto, SP - Brazil

^[6] Fundacao Fac Reg Med Hosp Base, Ambulatorio Oftalmol, Sao Jose Do Rio Preto, SP - Brazil

^[7] Hosp Crianca & Maternidade, Sao Jose Do Rio Preto, SP - Brazil

^[8] Fac Med Sao Jose do Rio Preto, Sao Jose Do Rio Preto, SP - Brazil

^[9] Univ Sao Paulo, Fac Med Vet & Zootecnia, Sao Paulo, SP - Brazil

^[10] Fac Med Marilia, Dept Oftalmol, Marilia, SP - Brazil

^[11] Univ Sao Paulo, Dept Oftalmol, Ribeirao Preto, SP - Brazil

• Document type: Journal article
• Source: Brazilian Journal of Infectious Diseases; v. 21, n. 6, p. 638-647, NOV-DEC 2017.
• Web of Science Citations: 5

Abstract

ABSTRACT Symptomatic forms of toxoplasmosis are a serious public health problem and occur in around 10-20% of the infected people. Aiming to improve the molecular diagnosis of symptomatic toxoplasmosis in Brazilian patients, this study evaluated the performance of real time PCR testing two primer sets (B1 and REP-529) in detecting Toxoplasma gondii DNA. The methodology was assayed in 807 clinical samples with known clinical diagnosis, ELISA, and conventional PCR results in a 9-year period. All samples were from patients with clinical suspicion of several features of toxoplasmosis. According to the minimum detection limit curve (in CT), REP-529 had greater sensitivity to detect T. gondii DNA than B1. Both primer sets were retrospectively evaluated using 515 DNA from different clinical samples. The 122 patients without toxoplasmosis provided high specificity (REP-529, 99.2% and B1, 100%). From the 393 samples with positive ELISA, 146 had clinical diagnosis of toxoplasmosis and positive conventional PCR. REP-529 and B1 sensitivities were 95.9% and 83.6%, respectively. Comparison of REP-529 and B1 performances was further analyzed prospectively in 292 samples. Thus, from a total of 807 DNA analyzed, 217 (26.89%) had positive PCR with, at least one primer set and symptomatic toxoplasmosis confirmed by clinical diagnosis. REP-529 was positive in 97.23%, whereas B1 amplified only 78.80%. After comparing several samples in a Brazilian referral laboratory, this study concluded that REP-529 primer set had better performance than B1 one. These observations were based after using cases with defined clinical diagnosis, ELISA, and conventional PCR. (AU)

• FAPESP's process: 13/15879-8 - Toxoplasma gondii infection: serological and molecular characterization in patients attended in public health service (SUS)
• Grantee: Fernando Henrique Antunes Murata
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 13/25650-8 - Class II HLA genes (DRB1 and DQB1) as risck factors for ocular toxoplasmosis
• Grantee: Luiz Carlos de Mattos
• Support Opportunities: Regular Research Grants
• FAPESP's process: 13/10050-5 - Serological, molecular and images in patients with active lesions or acute relapse of ocular infection by Toxoplasma gondii: a study follow-up
• Grantee: Mariana Previato
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 14/09496-1 - New diagnostic strategies employed in symptomatic toxoplasmosis
• Grantee: Vera Lúcia Pereira Chioccola
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/01706-7 - Genotypic characterization of Toxoplasma gondii in samples of pregnant women in the northwest region of the state of São Paulo
• Grantee: Marina Neves Ferreira
• Support Opportunities: Scholarships in Brazil - Scientific Initiation