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

被引:28
|
作者
Teixeira, Leandro Emidio [1 ]
Kanunfre, Kelly Aparecida [1 ,2 ]
Shimokawa, Paulo Tadashi [1 ]
Targa, Lilia Spaleta [1 ]
Rodrigues, Jonatas Cristian [1 ,2 ]
Domingues, Wilson [1 ]
Yamamoto, Lidia [1 ]
Okay, Thelma Suely [1 ]
机构
[1] Univ Sao Paulo, Inst Med Trop Sao Paulo, Lab Soroepidemiol & Imunobiol, Sao Paulo, Brazil
[2] Univ Sao Paulo, Fac Med, Dept Doencas Infecciosas & Parasitarias, Lab Imunol, Sao Paulo, Brazil
基金
巴西圣保罗研究基金会;
关键词
Congenital toxoplasmosis; Congenital infection; Molecular diagnosis; PCR; Quantitative PCR; Real-time PCR; REAL-TIME PCR; GONDII; INFECTION; AF146527; CHILDREN; REPEAT;
D O I
10.1590/0037-8682-0095-2013
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
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%).
引用
收藏
页码:584 / 588
页数:5
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