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Braz. j. med. biol. res ; 49(10): e5303, 2016. tab, graf
Article Dans Anglais | LILACS | ID: lil-792526

Résumé

The shipment and storage conditions of clinical samples pose a major challenge to the detection accuracy of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Ureaplasma urealyticum (UU) when using quantitative real-time polymerase chain reaction (qRT-PCR). The aim of the present study was to explore the influence of storage time at 4°C on the DNA of these pathogens and its effect on their detection by qRT-PCR. CT, NG, and UU positive genital swabs from 70 patients were collected, and DNA of all samples were extracted and divided into eight aliquots. One aliquot was immediately analyzed with qRT-PCR to assess the initial pathogen load, whereas the remaining samples were stored at 4°C and analyzed after 1, 2, 3, 7, 14, 21, and 28 days. No significant differences in CT, NG, and UU DNA loads were observed between baseline (day 0) and the subsequent time points (days 1, 2, 3, 7, 14, 21, and 28) in any of the 70 samples. Although a slight increase in DNA levels was observed at day 28 compared to day 0, paired sample t-test results revealed no significant differences between the mean DNA levels at different time points following storage at 4°C (all P>0.05). Overall, the CT, UU, and NG DNA loads from all genital swab samples were stable at 4°C over a 28-day period.


Sujets)
Humains , Mâle , Femelle , Adulte , Adulte d'âge moyen , Jeune adulte , Chlamydia trachomatis/génétique , ADN bactérien/isolement et purification , Neisseria gonorrhoeae/génétique , Réaction de polymérisation en chaine en temps réel/méthodes , Manipulation d'échantillons , Ureaplasma urealyticum/génétique , Charge bactérienne , Chlamydia trachomatis/isolement et purification , Système génital/microbiologie , Neisseria gonorrhoeae/isolement et purification , Valeurs de référence , Facteurs temps , Ureaplasma urealyticum/isolement et purification
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