Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7556616 | Analytical Biochemistry | 2018 | 6 Pages |
Abstract
In this work, we employed real-time PCR analysis targeting tp0574 to investigate the effects of different processing procedures on the yield of T. pallidum DNA from blood to improve assay sensitivity. The T. pallidum DNA yields following red blood cell lysis pretreatment were 40.4 times greater from whole blood and 32.4 times greater from residual hematocytes than yields without pretreatment. For the simulated whole-blood experiments, the T. pallidum DNA yields from the lower layer were 2.8, 4.6, 7.3, 12.6, 15.24, 16.7, 65.1 and 73.1 times those from the upper layer following centrifugation at 500Ã, 1000Ã, 2000Ã, 4000Ã, 5000Ã, 7000Ã, 10,000à and 20,000â¯Ãâ¯g, respectively. However, the T. pallidum DNA yields from blood clots were only 1.0% at different centrifugal forces. The experiment with infected rabbit blood showed results similar to those mentioned above. In addition, sample processing time (within 48â¯h) and storage temperature (4â¯Â°C and 25â¯Â°C) did not affect T. pallidum DNA extraction efficiency. The T. pallidum DNA yield can be significantly improved by red blood cell lysis pretreatment and appropriate centrifugation. Furthermore, the T. pallidum DNA extraction yield is greater from whole blood or residual hematocytes from anti-coagulated blood than from plasma, serum or blood clots.
Related Topics
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Chemistry
Analytical Chemistry
Authors
Xiao-Zhen Zhu, Jin-Yi Fan, Dan Liu, Zheng-Xiang Gao, Kun Gao, Yong Lin, Man-Li Tong, Hui-Lin Zhang, Li-Li Liu, Tian-Ci Yang, Li-Rong Lin, Jian-Jun Niu,