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首页> 外文期刊>Brazilian Journal of Medical and Biological Research >Influence of storage time on DNA of Chlamydia trachomatis, Ureaplasma urealyticum , and Neisseria gonorrhoeae for accurate detection by quantitative real-time polymerase chain reaction
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Influence of storage time on DNA of Chlamydia trachomatis, Ureaplasma urealyticum , and Neisseria gonorrhoeae for accurate detection by quantitative real-time polymerase chain reaction

机译：贮藏时间对沙眼衣原体，解脲脲原体和淋病奈瑟氏菌DNA的影响，可通过定量实时聚合酶链反应进行准确检测

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摘要

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.
机译：当使用定量实时聚合酶链反应（qRT-PCR）时，临床样品的运输和储存条件对沙眼衣原体（CT），淋病奈瑟氏球菌（NG）和解脲支原体（UU）的检测准确性提出了重大挑战。本研究的目的是探讨在4℃下储存时间对这些病原体DNA的影响及其对qRT-PCR检测的影响。收集了来自70例患者的CT，NG和UU阳性生殖器拭子，并提取了所有样本的DNA并分成8等份。立即用qRT-PCR分析一份样品，以评估初始病原体负荷，而其余样品则存储在4℃下，并在1、2、3、7、14、21和28天后进行分析。在70个样本中的任何一个样本中，在基线（第0天）与随后的时间点（第1、2、3、7、14、21和28天）之间，CT，NG和UU DNA负载均未观察到显着差异。尽管与第0天相比，在第28天观察到DNA水平略有增加，但成对的样本t检验结果显示，在4℃储存后，不同时间点的平均DNA水平之间没有显着差异（所有P> 0.05）。总体而言，所有生殖器拭子样品的CT，UU和NG DNA负荷在28天内稳定在4？C。

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