Increased detection of rotavirus using a real time reverse transcription-polymerase chain reaction (RT-PCR) assay in stool specimens from children with diarrhea. Pang, X. L, Lee, B., Boroumand, N., Leblanc, B., Preiksaitis, J. K, & Yu Ip, C. C Journal of Medical Virology, 72(3):496–501, March, 2004. Paper doi abstract bibtex Six-hundred and twenty-six stool specimens collected from children with diarrhea over a 12-month period were tested for rotavirus using a real time reverse transcription-polymerase chain reaction (RT-PCR) assay, a conventional nested PCR assay and by electron microscopy (EM). A fragment of 87 bp in a highly-conserved region of non-structural protein 3 (NSP3) in rotavirus genome was amplified by a single-step RT-PCR protocol in a closed-tube system. Rotavirus was detected in 123 samples (20%) with the real time RT-PCR assay, 113 samples (18%) with the nested-PCR assay, and 79 samples (13%) with EM. Using serial diluted nucleic acid extract, we compared the sensitivity of real time RT-PCR with conventional RT-PCR and conventional nested PCR assays. Real time RT-PCR was two to four logs more sensitive than the conventional assays. The reaction time required for the RT-PCR assay is about half the time required for the conventional nested-PCR. The real time RT-PCR assay is both simple and rapid with advantages including enhanced sensitivity and a lower risk for cross-contamination making it a useful tool for the detection of rotavirus in various situations including sporadic gastroenteritis, outbreaks, and environmental investigations. G(1) was the predominant type (89%), followed by G(2) (10%), and G(4) (1%). No rotavirus of G(3), G(8), and G(9) types were found. The peak season for rotavirus infection was January to May in northern Alberta.
@article{pang_increased_2004,
title = {Increased detection of rotavirus using a real time reverse transcription-polymerase chain reaction ({RT}-{PCR}) assay in stool specimens from children with diarrhea},
volume = {72},
issn = {0146-6615},
url = {http://www.ncbi.nlm.nih.gov/pubmed/14748075},
doi = {10.1002/jmv.20009},
abstract = {Six-hundred and twenty-six stool specimens collected from children with diarrhea over a 12-month period were tested for rotavirus using a real time reverse transcription-polymerase chain reaction (RT-PCR) assay, a conventional nested PCR assay and by electron microscopy (EM). A fragment of 87 bp in a highly-conserved region of non-structural protein 3 (NSP3) in rotavirus genome was amplified by a single-step RT-PCR protocol in a closed-tube system. Rotavirus was detected in 123 samples (20\%) with the real time RT-PCR assay, 113 samples (18\%) with the nested-PCR assay, and 79 samples (13\%) with EM. Using serial diluted nucleic acid extract, we compared the sensitivity of real time RT-PCR with conventional RT-PCR and conventional nested PCR assays. Real time RT-PCR was two to four logs more sensitive than the conventional assays. The reaction time required for the RT-PCR assay is about half the time required for the conventional nested-PCR. The real time RT-PCR assay is both simple and rapid with advantages including enhanced sensitivity and a lower risk for cross-contamination making it a useful tool for the detection of rotavirus in various situations including sporadic gastroenteritis, outbreaks, and environmental investigations. G(1) was the predominant type (89\%), followed by G(2) (10\%), and G(4) (1\%). No rotavirus of G(3), G(8), and G(9) types were found. The peak season for rotavirus infection was January to May in northern Alberta.},
number = {3},
urldate = {2011-11-01},
journal = {Journal of Medical Virology},
author = {Pang, Xiaoli L and Lee, Bonita and Boroumand, Nasim and Leblanc, Barbara and Preiksaitis, Jutta K and Yu Ip, Charlotte C},
month = mar,
year = {2004},
pmid = {14748075},
keywords = {\#duplicates},
pages = {496--501},
}
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A fragment of 87 bp in a highly-conserved region of non-structural protein 3 (NSP3) in rotavirus genome was amplified by a single-step RT-PCR protocol in a closed-tube system. Rotavirus was detected in 123 samples (20%) with the real time RT-PCR assay, 113 samples (18%) with the nested-PCR assay, and 79 samples (13%) with EM. Using serial diluted nucleic acid extract, we compared the sensitivity of real time RT-PCR with conventional RT-PCR and conventional nested PCR assays. Real time RT-PCR was two to four logs more sensitive than the conventional assays. The reaction time required for the RT-PCR assay is about half the time required for the conventional nested-PCR. The real time RT-PCR assay is both simple and rapid with advantages including enhanced sensitivity and a lower risk for cross-contamination making it a useful tool for the detection of rotavirus in various situations including sporadic gastroenteritis, outbreaks, and environmental investigations. G(1) was the predominant type (89%), followed by G(2) (10%), and G(4) (1%). No rotavirus of G(3), G(8), and G(9) types were found. The peak season for rotavirus infection was January to May in northern Alberta.","number":"3","urldate":"2011-11-01","journal":"Journal of Medical Virology","author":[{"propositions":[],"lastnames":["Pang"],"firstnames":["Xiaoli","L"],"suffixes":[]},{"propositions":[],"lastnames":["Lee"],"firstnames":["Bonita"],"suffixes":[]},{"propositions":[],"lastnames":["Boroumand"],"firstnames":["Nasim"],"suffixes":[]},{"propositions":[],"lastnames":["Leblanc"],"firstnames":["Barbara"],"suffixes":[]},{"propositions":[],"lastnames":["Preiksaitis"],"firstnames":["Jutta","K"],"suffixes":[]},{"propositions":[],"lastnames":["Yu","Ip"],"firstnames":["Charlotte","C"],"suffixes":[]}],"month":"March","year":"2004","pmid":"14748075","keywords":"#duplicates","pages":"496–501","bibtex":"@article{pang_increased_2004,\n\ttitle = {Increased detection of rotavirus using a real time reverse transcription-polymerase chain reaction ({RT}-{PCR}) assay in stool specimens from children with diarrhea},\n\tvolume = {72},\n\tissn = {0146-6615},\n\turl = {http://www.ncbi.nlm.nih.gov/pubmed/14748075},\n\tdoi = {10.1002/jmv.20009},\n\tabstract = {Six-hundred and twenty-six stool specimens collected from children with diarrhea over a 12-month period were tested for rotavirus using a real time reverse transcription-polymerase chain reaction (RT-PCR) assay, a conventional nested PCR assay and by electron microscopy (EM). A fragment of 87 bp in a highly-conserved region of non-structural protein 3 (NSP3) in rotavirus genome was amplified by a single-step RT-PCR protocol in a closed-tube system. Rotavirus was detected in 123 samples (20\\%) with the real time RT-PCR assay, 113 samples (18\\%) with the nested-PCR assay, and 79 samples (13\\%) with EM. Using serial diluted nucleic acid extract, we compared the sensitivity of real time RT-PCR with conventional RT-PCR and conventional nested PCR assays. Real time RT-PCR was two to four logs more sensitive than the conventional assays. The reaction time required for the RT-PCR assay is about half the time required for the conventional nested-PCR. The real time RT-PCR assay is both simple and rapid with advantages including enhanced sensitivity and a lower risk for cross-contamination making it a useful tool for the detection of rotavirus in various situations including sporadic gastroenteritis, outbreaks, and environmental investigations. G(1) was the predominant type (89\\%), followed by G(2) (10\\%), and G(4) (1\\%). No rotavirus of G(3), G(8), and G(9) types were found. The peak season for rotavirus infection was January to May in northern Alberta.},\n\tnumber = {3},\n\turldate = {2011-11-01},\n\tjournal = {Journal of Medical Virology},\n\tauthor = {Pang, Xiaoli L and Lee, Bonita and Boroumand, Nasim and Leblanc, Barbara and Preiksaitis, Jutta K and Yu Ip, Charlotte C},\n\tmonth = mar,\n\tyear = {2004},\n\tpmid = {14748075},\n\tkeywords = {\\#duplicates},\n\tpages = {496--501},\n}\n\n","author_short":["Pang, X. L","Lee, B.","Boroumand, N.","Leblanc, B.","Preiksaitis, J. K","Yu Ip, C. 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