Comparison of culture and real-time polymerase chain reaction methods for detection of Mycoplasma hominis in amniotic fluids samples.
Niger J Clin Pract
; 21(9): 1127-1131, 2018 Sep.
Article
em En
| MEDLINE
| ID: mdl-30156196
ABSTRACT
BACKGROUND:
Mycoplasma hominis is often present in the amniotic cavity with microbial invasion associated with spontaneous preterm labor. Conventional culture method is the gold standard for detection of Mycoplasmas, but real-time polymerase chain reaction (real-time PCR) has revolutionized the diagnosis of M. hominis.OBJECTIVE:
The purpose of this study is the comparison of the culture methodology with real-time PCR for the detection of M. hominis in amniotic fluid samples.METHODS:
Amniotic fluid samples were collected from 65 pregnant women (age range 25-45 years) previously followed at an infertility clinic. They were collected by transabdominal genetic amniocentesis during 16-21 weeks of gestation. Amniotic fluids were inoculated in SP4 broth for 48-72 h, and after becoming alkaline, culture suspension was spread on A7 agar plate for 1 week till the typical colonies seen in "fried-egg" morphology under stereomicroscope. DNA was extracted using a QIAGEN Mini DNA kit. The real-time-PCR was performed using Rotor-Gene Q Real-time PCR instrument. A melting-curve analysis was also performed. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were measured by real-time PCR by taking culture as gold standard.RESULTS:
Sixty-five women in 16-21 weeks of gestation, with a mean age of 33 ± 5.06 years, were enrolled into this study. M. hominis detected by culture and real-time PCR assay was 72% (47/65) and 69% (45/65), respectively. 66% (43/65) specimens were positive by both methods. Real-time PCR sensitivity was 91.5%, specificity 88.9%, PPV 95.6%, and NPV 80%.CONCLUSION:
Rapid detection of Mycoplasmas causing maternal complications such as neonatal infections and preterm labor in pregnancy by real-time PCR may be important and necessary. The high sensitivity and shorter time requirement of real-time PCR support its further development for diagnosis of Mycoplasma infections.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Adult
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Female
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Humans
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Middle aged
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Newborn
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Pregnancy
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article