Comparison of culture and real-time polymerase chain reaction methods for detection of Mycoplasma hominis in amniotic fluids samples

Nigerian Journal of Clinical Practice
F KeskinA Agacfidan

Abstract

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. 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. 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. Sixty-f...Continue Reading

Citations

Jul 28, 2020·The Canadian Journal of Infectious Diseases & Medical Microbiology = Journal Canadien Des Maladies Infectieuses Et De La Microbiologie Médicale·Xin-Ru MaoXiao-Mao Yin

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