Development of a droplet digital PCR method for the detection of Ureaplasma urealyticum
Background: Human infection with Ureaplasma urealyticum(UU) is mainly manifested as non-gonococcal urethritis, where it can lead to cervicitis, premature rupture of membranes and abortion in women, as well as infertility in males, which becomes a major problem in clinical diagnosis and treatment. At...
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Elsevier
2025-01-01
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| Series: | Practical Laboratory Medicine |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352551724000891 |
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| author | Yong-Zhuo Zhou Yun-Hu Zhao Yan-Lan Chen Hua Luo Yu-lin Zhou Bing Gu Wei-Zhen Fang Chao-Hui Duan Xu-Guang Guo |
| author_facet | Yong-Zhuo Zhou Yun-Hu Zhao Yan-Lan Chen Hua Luo Yu-lin Zhou Bing Gu Wei-Zhen Fang Chao-Hui Duan Xu-Guang Guo |
| author_sort | Yong-Zhuo Zhou |
| collection | DOAJ |
| description | Background: Human infection with Ureaplasma urealyticum(UU) is mainly manifested as non-gonococcal urethritis, where it can lead to cervicitis, premature rupture of membranes and abortion in women, as well as infertility in males, which becomes a major problem in clinical diagnosis and treatment. At present, real-time fluorescence quantitative PCR and culture are the two main methods for detecting UU. The real-time fluorescence quantitative PCR method is cumbersome and cannot accomplish absolute quantification on nucleic acids, while the cultivation method has limitations such as low sensitivity and being time-consuming. The aim of this study is to establish a more rapid and accurate droplet digital PCR(ddPCR) method for the detection of UU. Methods: Primers were designed for the ParC gene of UU. Nucleic acids from a standard strain of UU were extracted. Specificity, sensitivity, and repeatability detection was performed using ddPCR, and the detection performance of ddPCR was evaluated. Results: The detection process could be completed in 92 min. It has a high sensitivity of up to 3.8 pg/μL. With a high specificity, no positive microdrop were detected in eight negative control pathogens in this experiment. In addition, ddPCR detection of UU has good repeatability, and the calculated CV is 2.1 %. Conclusion: Our data indicated that ddPCR detection technology has the characteristics of absolute quantification, high stability, high specificity and high sensitivity of UU. It can promote the accurate detection of UU, providing a more scientific basis for clinical diagnosis and treatment. |
| format | Article |
| id | doaj-art-b4c7a1061fc74b3ca0796ee93b1cd498 |
| institution | Kabale University |
| issn | 2352-5517 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Practical Laboratory Medicine |
| spelling | doaj-art-b4c7a1061fc74b3ca0796ee93b1cd4982025-01-17T04:49:37ZengElsevierPractical Laboratory Medicine2352-55172025-01-0143e00443Development of a droplet digital PCR method for the detection of Ureaplasma urealyticumYong-Zhuo Zhou0Yun-Hu Zhao1Yan-Lan Chen2Hua Luo3Yu-lin Zhou4Bing Gu5Wei-Zhen Fang6Chao-Hui Duan7Xu-Guang Guo8Laboratory of Clinical, The Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, ChinaDepartment of Laboratory Medicine, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, ChinaGuangzhou Xiaoqi Biotechnology Co., LTD, Guangzhou, 510699, ChinaLaboratory of Clinical, The Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, ChinaDepartment of Clinical Laboratory Medicine, Guangdong Provincial Key Laboratory of Major Obstetric Diseases, Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510150, ChinaDepartment of Laboratory Medicine, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, ChinaLaboratory of Clinical, The Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, China; Corresponding author.Laboratory of Clinical, The Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, China; Corresponding author.Department of Clinical Laboratory Medicine, Guangdong Provincial Key Laboratory of Major Obstetric Diseases, Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510150, China; Department of Clinical Medicine, The Third Clinical School of Guangzhou Medical University, Guangzhou, 511436, China; Guangzhou Key Laboratory for Clinical Rapid Diagnosis and Early Warning of Infectious Diseases, King Med School of Laboratory Medicine, Guangzhou Medical University, Guangzhou, 510000, China; Corresponding author. Department of Clinical Laboratory Medicine, Guangdong Provincial Key Laboratory of Major Obstetric Diseases, Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510150, China.Background: Human infection with Ureaplasma urealyticum(UU) is mainly manifested as non-gonococcal urethritis, where it can lead to cervicitis, premature rupture of membranes and abortion in women, as well as infertility in males, which becomes a major problem in clinical diagnosis and treatment. At present, real-time fluorescence quantitative PCR and culture are the two main methods for detecting UU. The real-time fluorescence quantitative PCR method is cumbersome and cannot accomplish absolute quantification on nucleic acids, while the cultivation method has limitations such as low sensitivity and being time-consuming. The aim of this study is to establish a more rapid and accurate droplet digital PCR(ddPCR) method for the detection of UU. Methods: Primers were designed for the ParC gene of UU. Nucleic acids from a standard strain of UU were extracted. Specificity, sensitivity, and repeatability detection was performed using ddPCR, and the detection performance of ddPCR was evaluated. Results: The detection process could be completed in 92 min. It has a high sensitivity of up to 3.8 pg/μL. With a high specificity, no positive microdrop were detected in eight negative control pathogens in this experiment. In addition, ddPCR detection of UU has good repeatability, and the calculated CV is 2.1 %. Conclusion: Our data indicated that ddPCR detection technology has the characteristics of absolute quantification, high stability, high specificity and high sensitivity of UU. It can promote the accurate detection of UU, providing a more scientific basis for clinical diagnosis and treatment.http://www.sciencedirect.com/science/article/pii/S2352551724000891Droplet digital PCRUreaplasma urealyticumQuantitative testing |
| spellingShingle | Yong-Zhuo Zhou Yun-Hu Zhao Yan-Lan Chen Hua Luo Yu-lin Zhou Bing Gu Wei-Zhen Fang Chao-Hui Duan Xu-Guang Guo Development of a droplet digital PCR method for the detection of Ureaplasma urealyticum Practical Laboratory Medicine Droplet digital PCR Ureaplasma urealyticum Quantitative testing |
| title | Development of a droplet digital PCR method for the detection of Ureaplasma urealyticum |
| title_full | Development of a droplet digital PCR method for the detection of Ureaplasma urealyticum |
| title_fullStr | Development of a droplet digital PCR method for the detection of Ureaplasma urealyticum |
| title_full_unstemmed | Development of a droplet digital PCR method for the detection of Ureaplasma urealyticum |
| title_short | Development of a droplet digital PCR method for the detection of Ureaplasma urealyticum |
| title_sort | development of a droplet digital pcr method for the detection of ureaplasma urealyticum |
| topic | Droplet digital PCR Ureaplasma urealyticum Quantitative testing |
| url | http://www.sciencedirect.com/science/article/pii/S2352551724000891 |
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