ABSTRACT
<p><b>OBJECTIVE</b>To study the molecular characteristics of Noroviruses causing outbreaks of acute gastroenteritis in Huzhou.</p><p><b>METHODS</b>From 2008 to 2010, total 119 fecal specimens collected from outbreaks of acute gastroenteritis were tested for Norovirus. Partial sequence of RNA dependent RNA polymerase (RdRp) of the positive samples were amplified by RT-PCR, then the PCR production were purified, sequenced and put into phylogenetic analysis.</p><p><b>RESULTS</b>50 of 119 specimens were positive for Norovirus by real-time RT-PCR. Out of those 50 Norovirus positive specimens, 9 were Norovirus Genogroup I (GI) positive, 35 were Norovirus Genogroup II (GII) positive, 6 was both Norovirus GI and GII positive. 12 PCR products for RdRp were selected for further studies on sequencing. Phylogenetic analysis revealed that the 5 GI norovirus isolates were belonged to genotype GI/2 and GI/3. Of the 7 GII norovirus isolates, 6 were belonged to genotype GII/4, 1 was belonged to genotype Glib.</p><p><b>CONCLUSION</b>Norovirus is a major cause of outbreaks of acute gastroenteritis in Huzhou and the epidemic strains of norovirus isolated from Huzhou had a high degree of genetic diversity.</p>
Subject(s)
Female , Humans , Male , Acute Disease , China , Epidemiology , Disease Outbreaks , Gastroenteritis , Epidemiology , Genetic Variation , Norovirus , Classification , Genetics , Phylogeny , RNA-Dependent RNA Polymerase , Genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
OBJECTIVE@#The aim of the present study was to establish a rapid and robust assay used to simultaneously genotype SNPs by the single nucleotide primer extension (minisequencing) with the SNaPshot Kit and obtain the population genetic data in Chinese population in Sichuan. The analysis of single nucleotide polymorphisms (SNPs) is a promising application in forensic casework.@*METHODS@#12 Y-SNPs, which were SRY2627, SRY1532, M13, M20, SRY8299, Tat, M69, M9, 92R7, M17, M19 and M112, were multiple amplificated and the PCR products were pooled, Purified, and then used as templates for the minisequencing reaction with the commol/Lercially available SNaPshot Kit. Then the products of minisequencing reaction were detected by capillary elcetrophoresis.@*RESULTS@#78 genomic DNA individual samples from Sichuan, China and 5 semen stain samples from sexal criminal scene were analyzed and two haplotypes could be identified.@*CONCLUSION@#A rapid method has been established to analyze the 12 Y-SNPs by multiplex PCR and minisequencing. It can be applied in forensic casework successfully.
Subject(s)
Humans , Male , Base Sequence , Bone and Bones/chemistry , China/ethnology , Chromosomes, Human, Y , DNA/analysis , DNA Fingerprinting/methods , DNA Primers , Electrophoresis, Capillary , Forensic Medicine , Gene Frequency , Genetic Markers , Genetics, Population , Polymerase Chain Reaction/methods , Polymorphism, Single Nucleotide , Sequence Analysis, DNA/methods , Sex Offenses , Tandem Repeat SequencesABSTRACT
24 hours) and versus control group(