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

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 an...

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Published in:	The Brazilian journal of infectious diseases Vol. 21; no. 6; pp. 638 - 647
Main Authors:	Camilo, Lilian Muniz, Pereira-Chioccola, Vera Lucia, Gava, Ricardo, Meira-Strejevitch, Cristina da Silva, Vidal, Jose Ernesto, Brandão de Mattos, Cinara Cássia, Frederico, Fábio Batista, De Mattos, Luiz Carlos, Spegiorin, Lígia Cosentino Junqueira Franco, Murata, Fernando Henrique Antunes, Ferreira, Marina Neves, Barbosa, Deusenia Machado Ulisses, Gonçalves, Fausto da Silva, Dias, Cristiane Moraes, Catelan, Marcia Wakai, Siqueira, Rubens Camargo, Previato, Mariana, Barbosa, Amanda Pires, Cavallini, Danilo
Format:	Journal Article
Language:	English
Published:	Brazil Elsevier Editora Ltda 01-11-2017 Contexto Elsevier Brazilian Society of Infectious Diseases
Subjects:	Diagnosis DNA Primers - genetics DNA, Protozoan - genetics Humans INFECTIOUS DISEASES Molecular diagnostic techniques Original Prospective Studies Real-Time Polymerase Chain Reaction Retrospective Studies Sensitivity and Specificity Symptomatic toxoplasmosis Toxoplasma - genetics Toxoplasma gondii Toxoplasmosis Toxoplasmosis - classification Toxoplasmosis - diagnosis Symptomatic toxoplasmosis Toxoplasma gondii Diagnosis Real-time polymerase chain reaction
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1413-8670 1678-4391 1678-4391
DOI:	10.1016/j.bjid.2017.07.003