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.
Increased detection of rotavirus using a real time reverse transcription-polymerase chain reaction (RT-PCR) assay in stool specimens from children with diarrhea [link]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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