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Braz J Med Biol Res ; 49(10): e5303, 2016 Aug 25.
Article in English | MEDLINE | ID: mdl-27580005

ABSTRACT

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.


Subject(s)
Chlamydia trachomatis/genetics , DNA, Bacterial/isolation & purification , Neisseria gonorrhoeae/genetics , Real-Time Polymerase Chain Reaction/methods , Specimen Handling , Ureaplasma urealyticum/genetics , Adult , Bacterial Load , Chlamydia trachomatis/isolation & purification , Female , Genitalia/microbiology , Humans , Male , Middle Aged , Neisseria gonorrhoeae/isolation & purification , Reference Values , Time Factors , Ureaplasma urealyticum/isolation & purification , Young Adult
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