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- Title
Elimination of human cytomegalovirus DNA degradation in urine.
- Authors
Guo, Xiaoyi; Wang, Han; You, Ruilan; Tang, Jiabao; Wang, Jin; Song, Qiaoqiao; Huang, Yue; Zhang, Xuejie; Zhang, Shiyin; Li, Tingdong; Ge, Shengxiang; Wu, Ting; Zhang, Jun; Xia, Ningshao
- Abstract
Congenital cytomegalovirus infection (cCMVi) is an important cause of sensorineural hearing loss in newborns. Detection of human cytomegalovirus (HCMV) DNA in urine has been used to screen for cCMVi in newborns. However, the matrix effect of urine on HCMV DNA detection is unclear. To evaluate the matrix effect of urine on HCMV DNA detection and optimize the sample process strategy to eliminate or minimize the impact of urine on HCMV DNA detection, DNA in spiked samples was extracted using different DNA extraction methods, and urine samples that could inhibit HCMV DNA detection were mixed to evaluate the inhibitory substances, inhibitory mechanism, and elimination of the inhibitory effect. The optimal urine sample process strategy was evaluated using 42 adult female urine samples and 42 newborn urine samples spiked with HCMV. Some urine samples were found to inhibit HCMV DNA detection due to DNA degradation. The addition of ≥5 mM EDTA to the urine before extraction eliminated the inhibitory effect of urine and did not affect the detection results of urine exhibiting no inhibition. Of the 42 adult female and 42 newborn urine samples, four and two samples, respectively, could inhibit HCMV DNA detection. However, the inhibitory effects of these six urine samples were eliminated after the addition of EDTA. The collective results indicate that the addition of EDTA can completely eliminate the impact of inhibitors present in urine on HCMV DNA extraction and improve the detection of HCMV in urine. Highlights: Some urine can degrade HCMV DNA and inhibit the DNA detection.Small soluble molecules can directly or indirectly inhibit the detection of HCMV DNA.EDTA can fully eliminate the inhibitory effect of urine on HCMV DNA detection.
- Subjects
HUMAN DNA; ETHYLENEDIAMINETETRAACETIC acid; HUMAN cytomegalovirus; NUCLEIC acid isolation methods; URINE; CYTOMEGALOVIRUS diseases; DISEASE eradication
- Publication
Journal of Medical Virology, 2021, Vol 93, Issue 8, p5033
- ISSN
0146-6615
- Publication type
Article
- DOI
10.1002/jmv.27035