The performance of four molecular methods for the laboratory diagnosis of congenital toxoplasmosis in amniotic fluid samples

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dc.contributor Sistema FMUSP-HC: Faculdade de Medicina da Universidade de São Paulo (FMUSP) e Hospital das Clínicas da FMUSP
dc.contributor.author TEIXEIRA, Leandro Emidio
KANUNFRE, Kelly Aparecida FMUSP-HC
SHIMOKAWA, Paulo Tadashi FMUSP-HC
TARGA, Lilia Spaleta FMUSP-HC
RODRIGUES, Jonatas Cristian FMUSP-HC
DOMINGUES, Wilson
YAMAMOTO, Lidia FMUSP-HC
OKAY, Thelma Suely FMUSP-HC
dc.date.issued 2013
dc.identifier.citation REVISTA DA SOCIEDADE BRASILEIRA DE MEDICINA TROPICAL, v.46, n.5, p.584-588, 2013
dc.identifier.issn 0037-8682
dc.identifier.uri http://observatorio.fm.usp.br/handle/OPI/4178
dc.description.abstract Introduction: Toxoplasmosis may be life-threatening in fetuses and in immune-deficient patients. Conventional laboratory diagnosis of toxoplasmosis is based on the presence of IgM and IgG anti-Toxoplasma gondii antibodies; however, molecular techniques have emerged as alternative tools due to their increased sensitivity. The aim of this study was to compare the performance of 4 PCR-based methods for the laboratory diagnosis of toxoplasmosis. One hundred pregnant women who seroconverted during pregnancy were included in the study. The definition of cases was based on a 12-month follow-up of the infants. Methods: Amniotic fluid samples were submitted to DNA extraction and amplification by the following 4 Toxoplasma techniques performed with parasite B1 gene primers: conventional PCR, nested-PCR, multiplex-nested-PCR, and real-time PCR. Seven parameters were analyzed, sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (PLR), negative likelihood ratio (NLR) and efficiency (Ef). Results: Fifty-nine of the 100 infants had toxoplasmosis; 42 (71.2%) had IgM antibodies at birth but were asymptomatic, and the remaining 17 cases had non-detectable IgM antibodies but high IgG antibody titers that were associated with retinochoroiditis in 8 (13.5%) cases, abnormal cranial ultrasound in 5 (8.5%) cases, and signs/symptoms suggestive of infection in 4 (6.8%) cases. The conventional PCR assay detected 50 cases (9 false-negatives), nested-PCR detected 58 cases (1 false-negative and 4 false-positives), multiplex-nested-PCR detected 57 cases (2 false-negatives), and real-time-PCR detected 58 cases (1 false-negative). Conclusions: The real-time PCR assay was the best-performing technique based on the parameters of Se (98.3%), Sp (100%), PPV (100%), NPV (97.6%), PLR (co), NLR (0.017), and Ef (99%).
dc.description.sponsorship · FAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo) [2010/15022-1]
· CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico) [2011-0/471479]
dc.language.iso eng
dc.publisher SOC BRASILEIRA MEDICINA TROPICAL
dc.relation.ispartof Revista da Sociedade Brasileira de Medicina Tropical
dc.rights openAccess
dc.subject Congenital toxoplasmosis; Congenital infection; Molecular diagnosis; PCR; Quantitative PCR
dc.subject.other real-time pcr; gondii; infection; af146527; children; repeat
dc.title The performance of four molecular methods for the laboratory diagnosis of congenital toxoplasmosis in amniotic fluid samples
dc.type article
dc.rights.holder Copyright SOC BRASILEIRA MEDICINA TROPICAL
dc.description.group LIM/48
dc.description.group LIM/36
dc.identifier.doi 10.1590/0037-8682-0095-2013
dc.identifier.pmid 24409481
dc.type.category original article
dc.type.version publishedVersion
hcfmusp.author KANUNFRE, Kelly Aparecida:HC:LIM/48
hcfmusp.author SHIMOKAWA, Paulo Tadashi:IMT:
hcfmusp.author TARGA, Lilia Spaleta:IMT:
hcfmusp.author RODRIGUES, Jonatas Cristian:HC:LIM/48
hcfmusp.author YAMAMOTO, Lidia:IMT:
hcfmusp.author OKAY, Thelma Suely:FM:MPE
hcfmusp.author.external · TEIXEIRA, Leandro Emidio:Univ Sao Paulo, Inst Med Trop Sao Paulo, Lab Soroepidemiol & Imunobiol, Sao Paulo, Brazil
· DOMINGUES, Wilson:Univ Sao Paulo, Inst Med Trop Sao Paulo, Lab Soroepidemiol & Imunobiol, Sao Paulo, Brazil
hcfmusp.origem.id 2-s2.0-84888156919
hcfmusp.origem.id WOS:000327672500009
hcfmusp.origem.id SCIELO:S0037-86822013000500584
hcfmusp.publisher.city BRASILIA
hcfmusp.publisher.country BRAZIL
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dc.description.index MEDLINE
hcfmusp.citation.scopus 26
hcfmusp.affiliation.country Brasil
hcfmusp.scopus.lastupdate 2021-09-17


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