Abstract:
:The detection of the human RNA viruses, calicivirus and astrovirus, requires high sensitivity and broad reactivity. A novel single-tube nested reverse transcription-PCR (RT-PCR) method is described here, in which all of the required reagents are included in the one tube; however, those required for the nested amplification are separated in a "hanging drop" in the cap to be introduced by centrifugation after the RT and first-round cDNA amplification steps. Broad reactivity was obtained by using primer cocktails covering the published sequence variation in the primer targets. The method was evaluated with clinical fecal samples from outbreak and sporadic cases. Norwalk-like virus types 1 and 2 and rotavirus were the causal agents in 10 of 12 outbreaks. A viral agent was detected in 44% of 197 samples from sporadic infections in patients presenting to community health centers and a children's hospital. Interestingly, whereas rotavirus was more common than astrovirus in patients presenting to the hospital (33 and 7.6%, respectively), the reverse was true for patients presenting to community health centers (4.2 and 34%, respectively).
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Ratcliff RM,Doherty JC,Higgins GDdoi
10.1128/jcm.40.11.4091-4099.2002subject
Has Abstractpub_date
2002-11-01 00:00:00pages
4091-9issue
11eissn
0095-1137issn
1098-660Xjournal_volume
40pub_type
杂志文章abstract::We studied nine solid and two liquid media for their suitability to select Aeromonas and Plesiomonas spp. from human stools, using artificially contaminated samples as well as 254 samples from outpatients with and without diarrhea. Media with optimal sensitivity and specificity for Aeromonas spp. were alkaline peptone...
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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