Quantitative real-time polymerase chain reaction for the accurate detection of Toxoplasma gondii in amniotic fluid.
Diagn Microbiol Infect Dis
; 63(1): 10-5, 2009 Jan.
Article
em En
| MEDLINE
| ID: mdl-18990529
ABSTRACT
Infection with Toxoplasma gondii during pregnancy is often asymptomatic and may cause severe fetal damage. A quantitative TaqMan minor groove binder real-time polymerase chain reaction (PCR) assay was developed for the specific and sensitive detection of the previously described 529-bp repeat element occurring up to 200 to 300 times in T. gondii genome. The qualitative and quantitative detection limits determined were 6 and 20 marker copies (1/30 to 1/50 of 1 parasite) per PCR, respectively. In addition to standard PCR cycling conditions, 3 different fast PCR protocols were evaluated to minimize run time. A higher variability but no loss of specificity was observed. For the evaluation of clinical applicability, a total of 135 amniotic fluid samples were analyzed targeting both 529-bp and B1 gene. The sensitivity and specificity were 88.0% and 100.0% for B1, and 100.0% and 98.2% for 529-bp PCR assay (positive predictive value and negative predictive value 100.0% and 97.4%, and 92.6% and 100.0%, respectively). Our results demonstrated an increased sensitivity of the 529-bp PCR assay even in a faster protocol.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Toxoplasma
/
Toxoplasmose Congênita
/
Reação em Cadeia da Polimerase
/
Líquido Amniótico
Tipo de estudo:
Diagnostic_studies
/
Guideline
/
Prognostic_studies
/
Qualitative_research
Limite:
Animals
/
Female
/
Humans
/
Pregnancy
Idioma:
En
Revista:
Diagn Microbiol Infect Dis
Ano de publicação:
2009
Tipo de documento:
Article
País de afiliação:
Áustria