Comparison of two nucleic acid extraction methods in detecting influenza

doi:10.3877/cma.j.issn.1674-1358.2017.05.010

Abstract

Abstract:

Objective

To compare and evaluate the efficacy of the two different nucleic acid extraction methods for influenza determination.

Methods

There were 592 clinical suspected cases pharynx samples were collected. There were two methods were used to extract nucleic acid, such as column extraction and Trizol manual method. After extraction, RNA Nucleic acid were used in the RT-PCR and test with the electrophoresis. When there was a 564 bp marker positive, the sample would be sent to be tested.

Results

From December 2014 to April 2015, the influenza A was the main epidemic in Beijing; and less influenza B in Beijing. There were 66 (11.1%) Trizol positive while 222 (37.5%) positive for the column extraction.

Conclusions

The main epidemic influenza virus is the influenza A in Beijing. Column extraction is more efficient than the Trizol manual method. Use the column extraction, the diagnosis correction could be improved.

Key words: Influenza, Column extraction, Trizol manual method, Nucleic acid extraction

Ya’nan Liu, Jun Li, Wenhao Hua, Shujing Song, Xiaoliang Ma, Jianping Dong, Rui Song. Comparison of two nucleic acid extraction methods in detecting influenza[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2017, 11(05): 474-478.

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References 25

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试剂	剂量（μl）
Prime Seript 1 Step Enzyme Mix（酶）	0.4
2× 1 Step Buffer	5
Primer F1	0.3
Primer R1	0.3
RNase Free dH₂O	3
样本RNA	1

试剂	剂量（μl）
Prime Seript 1 Step Enzyme Mix（酶）	0.4
2× 1 Step Buffer	5
Primer F1	0.3
Primer R1	0.3
RNase Free dH₂O	3
样本RNA	1

试剂	剂量（μl）
Taq酶mix	15
Primer F2	1
Primer R2	1
RNase Free dH₂O	11
第一轮PCR扩增产物	2

试剂	剂量（μl）
Taq酶mix	15
Primer F2	1
Primer R2	1
RNase Free dH₂O	11
第一轮PCR扩增产物	2

方法	真阳性（例）	真阴性（例）	假阳性（例）	假阴性（例）	灵敏度（%）	特异度（%）
离心柱法	222	370	0	0	100.0	100.0
Trizol法	66	370	0	156	29.7	100.0

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