Author: Yu, Fei; Qiu, Ting; Zeng, Ying; Wang, Yiyin; Zheng, Shufa; Chen, Xiao; Chen, Yu
Title: Comparative Evaluation of Three Preprocessing Methods for Extraction and Detection of Influenza A Virus Nucleic Acids from Sputum Document date: 2018_3_2
ID: 0kt9b1ww_22
Snippet: Sufficient nucleic acid extraction and the removal of substances that inhibit amplification are critical factors influencing the detection of IAV by rRT-PCR (10). Comparison of 49 positive specimens in all of the three groups demonstrated that the Ct values of the DTT and PK groups were significantly lower and their nucleic acid concentration and A260/A280 ratio within 1.8-2.0 were higher than those of the NS group. This finding might be attribut.....
Document: Sufficient nucleic acid extraction and the removal of substances that inhibit amplification are critical factors influencing the detection of IAV by rRT-PCR (10). Comparison of 49 positive specimens in all of the three groups demonstrated that the Ct values of the DTT and PK groups were significantly lower and their nucleic acid concentration and A260/A280 ratio within 1.8-2.0 were higher than those of the NS group. This finding might be attributed to the inability of NS to dilute sticky sputum through violent vortex. Some cells cannot be released from sticky sputum and thus lead to inadequate specimen decomposition and RNA extract reduction. Meanwhile, high amounts of proteins in sticky liquid are retained and can influence PCR by disturbing the cell lysis of nucleic acid extraction that inhibits DNA polymerase activity.
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