PMID- 16687177
OWN - NLM
STAT- MEDLINE
DCOM- 20061031
LR  - 20061115
IS  - 0166-0934 (Print)
IS  - 0166-0934 (Linking)
VI  - 136
IP  - 1-2
DP  - 2006 Sep
TI  - Amplification of the entire genome of influenza A virus H1N1 and H3N2 subtypes by 
      reverse-transcription polymerase chain reaction.
PG  - 38-43
AB  - This study describes the development of a simple RT-PCR method to amplify the 
      whole genome of the influenza A virus based on the amplification of full-length 
      gene segments. Primers were designed based on the conserved regions of both the 
      5'-end and the 3'-end of each gene segment. After optimizing the duration and 
      temperature of denaturing, annealing, and extension, these primers could amplify 
      all of the full-length gene segments. To test the accuracy of the method, all 
      amplicons were subjected to DNA sequencing with an autosequencer. Eighteen 
      strains of influenza A virus which belonged to H1N1 or H3N2 subtypes were tested. 
      All eight segments of both subtypes were successfully amplified in all tested 
      strains. Using a newly developed reverse-transcriptase (RT), primers and PCR 
      running conditions, this study established a protocol to amplify the entire 
      genome of the influenza A virus. This method provides a tool which can be used 
      for the amplification of all genes of the H1N1 and H3N2 subtypes of influenza A 
      virus prior to analysis of their sequences, and to construct expression plasmids 
      for further study.
FAU - Chan, Chi-Ho
AU  - Chan CH
AD  - Department of Microbiology and Immunology, Chung Shan Medical University (CSMU), 
      110, Sec. No. 1, Chien-Kuo N. Road, Taichung 402, Taiwan, Republic of China. 
      chiho@csmu.edu.tw
FAU - Lin, Keh-Liang
AU  - Lin KL
FAU - Chan, You
AU  - Chan Y
FAU - Wang, Ya-Li
AU  - Wang YL
FAU - Chi, Yu-Tsai
AU  - Chi YT
FAU - Tu, Hsiao-Li
AU  - Tu HL
FAU - Shieh, Happy-K
AU  - Shieh HK
FAU - Liu, Wu-Tse
AU  - Liu WT
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20060509
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA Primers)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - DNA Primers
MH  - *Genome, Viral
MH  - Humans
MH  - Influenza A Virus, H1N1 Subtype/*genetics
MH  - Influenza A Virus, H3N2 Subtype/*genetics
MH  - RNA, Viral/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
EDAT- 2006/05/12 09:00
MHDA- 2006/11/01 09:00
CRDT- 2006/05/12 09:00
PHST- 2005/09/05 00:00 [received]
PHST- 2006/03/29 00:00 [revised]
PHST- 2006/03/30 00:00 [accepted]
PHST- 2006/05/12 09:00 [pubmed]
PHST- 2006/11/01 09:00 [medline]
PHST- 2006/05/12 09:00 [entrez]
AID - S0166-0934(06)00120-0 [pii]
AID - 10.1016/j.jviromet.2006.03.027 [doi]
PST - ppublish
SO  - J Virol Methods. 2006 Sep;136(1-2):38-43. doi: 10.1016/j.jviromet.2006.03.027. 
      Epub 2006 May 9.