1.Genetic characteristics of VP1 region of coxsackievirus A10 strains in Ningxia Hui Autonomous Region during 2013-2014.
Jiangtao MA ; Fang YUAN ; Hong LIN ; Hui CHEN ; Xuemin MA ; Jun ZHAN
Chinese Journal of Preventive Medicine 2015;49(12):1056-1060
OBJECTIVETo investigate the genetic characteristics of coxsackievirus A10(CV-A10) strains isolated from hand, foot and mouth disease (HFMD) cases in Ningxia province.
METHODSBased on the HFMD laboratory network surveillance system, 2 470 patients clinical specimens including 450 faeces and 2 020 throat swaps were collected from various regions people's hospital in Ningxia Hui Autonomous Region during January, 2013 to December, 2014. All specimens were isolated using rhabdomyosarcoma cells. VP1 regional gene of isolated strains was amplified by RT-PCR using degenerate primers and sequenced. Sequences were compared with the database of GenBank by the Blast algorithm to identify the enterovirus genotypes. All the CV-A10 strains were performed the homology and phylogenetic evolution analysis.
RESULTS450 specimens identified as non-EV-A71, non-CV-A16 enterovirus were collected and 36 CV-A10 strains were isolated, 6 strains were isolated in 2013 and 30 strains were isolated in 2014. The homology of nucleotides and amino acids among 36 CV-A10 strains were 90.6%-100.0% , and 90.2%-100.0%, respectively. Compared 36 strains with genotype A, B, C, D representative strains, it has the highest homology with the genotype C, the nucleotide and amino acids homogeneity were 90.2%-98.9% and 95.7%-99.7%. The phylogenetic tree showed 36 strains and genotype C representative strains located in the same evolutionary branch.
CONCLUSIONCV-A10 was one of the most common pathogen of HFMD in Ningxia Hui Autonomous Region. All CV-A10 strains belonged to genotype C and contained wide homology range.
China ; Enterovirus ; genetics ; Genotype ; Hand, Foot and Mouth Disease ; virology ; Humans ; Phylogeny ; Sequence Homology, Amino Acid ; Sequence Homology, Nucleic Acid
2.Nucleotide sequence and protein sequence analysis of GL-7-ACA acylase from Pseudomonas sp. 130.
Xiang MAO ; Jing ZHANG ; Yong LI ; Yu-Jiong HE ; En-Duo WANG ; Yun-Liu YANG ; Wei-Hong JIANG ; Guo-Ping ZHAO ; Jui-Sheng CHIAO
Chinese Journal of Biotechnology 2002;18(1):45-50
The nucleotide sequence and N-, C-terminal amino acid sequences of alpha,beta-subunit of glutaryl 7-ACA acylase C130 from Pseudomonas sp. 130 were determined. The alignment of the acylase C130 with the other acylases shows that it has high homology with the acylases from Pseudomonas sp. GK16 and C427, but low homology with the others. There is large difference in the N-terminal of alpha-subunit, while the N-terminal of beta-subunit has significant conservation.
Amino Acid Sequence
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Base Sequence
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DNA, Bacterial
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analysis
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Genes, Bacterial
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Molecular Sequence Data
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Penicillin Amidase
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genetics
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Pseudomonas
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enzymology
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genetics
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Sequence Homology, Amino Acid
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Sequence Homology, Nucleic Acid
3.Nucleotides sequence analysis of EV71 Chinese strain SHZH98 3C segment.
Ya-qing HE ; Fan YANG ; Liang-cheng LI ; Qi JIN
Chinese Journal of Experimental and Clinical Virology 2003;17(4):354-357
OBJECTIVETo study the EV71 Chinese strain SHZH98 and analyze its genetic evolution using 3c gene as index.
METHODSThe 3C gene cDNA of EV71 Chinese strain SHZH98 was amplified by PCR, the PCR product was sequenced.
RESULTSThe EV71 Chinese mainland strain SHZH98 3C segment was 549 bps in length. Comparison of nucleotide sequences from other enteroviruses which have been published, revealed a higher homology to strain MS, 78.7% at nucleotide level and 93.45% at deduced amino acid level. The homology to strain BrCr was 76.7% at nucleotide level and 89.1% at deduced amino acid level. Taiwan strains POLY,NCKU,TW2086,TW2272 shared a lower homology with Chinese mainland strain SHZH98, 74.0%, 73.8%, 71.9%, 69.8% at nucleotide level and 90.7%, 90.2%, 84.2%, 82.5% at deduced amino acid level. The genetic progress analysis revealed that EV71 Chinese mainland strain SHZH98 3C segment shares more homology with European and American strains than Taiwan strains.
CONCLUSIONThe non-structural protein of EV71 Chinese strains may have different evolutionary process from Taiwan strains.
Amino Acid Sequence ; Base Sequence ; Enterovirus ; genetics ; isolation & purification ; Genes, Viral ; Humans ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Amino Acid ; Sequence Homology, Nucleic Acid
4.Characterization of HA1 gene of influenza B virus circulated in 1990 through 2000 in China.
Jiming CHEN ; Yuanji GUO ; Junfeng GUO ; Jie DONG
Chinese Journal of Experimental and Clinical Virology 2002;16(3):278-280
OBJECTIVETo characterize HA1 gene of influenza B virus circulated in 1990 through 2000 in China.
METHODSViral RNA was extracted and transcribed into cDNA by reverse transcriptase and amplified by PCR. The product of PCR was purified and sequenced by ABI377. The sequence data were analyzed with epidemic records.
RESULTS1. Two major lineages of influenza B virus always circulated during the period of 1990-2000 in China; the Yamagata lineage was the main lineage, but in 1994 and 1997 the Victoria lineage was more active. 2. During 1992-2000 the Yamagata lineage evolved into two minor groups whose distance in HAI amino acid sequences was about 6%. 3. Large and non-reverse mutators led the development of influenza B epidemics in 1990-2000 in China. 4. Except for a few strains, there was little difference among the influenza B viruses of the same major lineages circulated in the same year in China.
CONCLUSIONSTwo major lineages of influenza B virus always circulated during the period from 1990-2000 in China,and the Yamagata lineage diverged into two minor groups in recent years. Exchanges of the lineages and the appearance of large non-reverse mutators possibly had important epidemic significance.
China ; epidemiology ; Genes, Viral ; genetics ; Influenza B virus ; classification ; genetics ; RNA, Viral ; genetics ; Sequence Analysis ; Sequence Homology, Amino Acid ; Sequence Homology, Nucleic Acid
6.Human DNA contains sequences homologous to the 5'-non-coding region of hepatits C virus: characterization with restriction endonucleases reveals individual varieties.
Chinese Medical Journal 2003;116(7):1092-1098
OBJECTIVETo investigate a 272 base pair section of the 5'-non-coding region of genomic DNA from the peripheral blood monounuclear cells of healthy hepatitis virus C (HCV)-negative human subjects (not patients).
METHODSThis sequence section bears interest because (1) it harbors several potential methylation (Cp-rich) sites, and (2) it represents the largest part of its internal ribosomal entry site. A pre-PCR digestion protocol was established making consistent use of four restriction endonucleases selected for certain features: SmaI, XmaCI, MspI, and HpaII are inhibited if methylation(s) are present at certain cytosines within their cutting sequences.
RESULTSThe suspected HCV-specific sequence was found in the DNA of each subject tested. The pre-PCR digestion assay reveals individual differences in their pattern of methylation, which may be due to possible epigenetic phenomena.
CONCLUSIONSThe results provide formal proof that these HCV-specific sequences are contained in the genomic or extra chromosomal target DNA, and probably belong to a new class of endogenous sequences.
DNA Methylation ; Hepacivirus ; genetics ; Humans ; Restriction Mapping ; Sequence Homology, Nucleic Acid
7.Genetic diversity of porcine sapoviruses from Lulong county in China.
Miao JIN ; Jie-Mei YU ; Hui-ying LI ; Qing ZHANG ; Shu-xian CUI ; Jing-yu TANG ; Zhao-jun DUAN
Chinese Journal of Virology 2010;26(3):255-259
Porcine sapoviruses (SaVs), which belong to the family Caliciviridae, have been considered potential zoonotic agents for human infection, and several cases have been reported in Asian countries. In this study, a total of 200 porcine fecal samples collected from Lulong county of China were tested. Among 200 samples, porcine sapoviruses were detected by RT-PCR in 17 samples (8.5%) showing their circulation in China. 14 out of 17 positive sapovirus strains were genetically related to the genogroup III (GIII) and were further divided into three different clusters or genotypes according to the phylogenetic analysis. In addition, the remaining three sapovirus strains belonged to GVII (one strain) and a potential novel genogroup (two strains) according to the phylogenetic analysis and the nucleotide identity and amino acid identity. These data suggested the genetic diversity of porcine sapoviruses in China.
Animals
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China
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Genetic Variation
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Genotype
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Phylogeny
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Sapovirus
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classification
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genetics
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Sequence Homology, Nucleic Acid
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Swine
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virology
8.Nucleotide sequencing of E/NS1 gene segment of dengue type 1 viruses isolated in Guangdong Province.
Kui ZHENG ; Limin JIANG ; Huiming LUO ; Jianfeng HE ; Xinmin DONG
Chinese Journal of Experimental and Clinical Virology 2002;16(4):382-384
OBJECTIVETo analyse the genetic relationship of local strains of dengue type 1 viruses isolated in different years and regions from Guangdong Province, and to explore the genetic links with strains of adjacent countries by comparing with the sequences of relevant strains in Genbank.
METHODSThe viral RNAs were extracted and used for one-step reverse transcriptase polymerase chain reaction (RT-PCR) to amplify the partial nucleotide fragments in E/NS1 gene junction which were then cloned into the plasmid pBluescript II SK for sequencing, the results were analysed by DNASTAR software.
RESULTSThe phylogenetic tree of the sequenced 14 strains of dengue type 1 viruses branches into two genotypic groups. The nucleotide sequences showed a maximal homologies of 99.2% with Indonesia strains, 100% with Philippines strains and 98.8% with Thailand strains.
CONCLUSIONSThe dengue type 1 viruses of Guangdong Province are closely related to the Philippines, Indonesia and Thailand strains, which may indicate the possibility of importation from those countries.
Base Sequence ; China ; Dengue Virus ; classification ; genetics ; isolation & purification ; Genes, Viral ; genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid
9.Identification of Swertia mussotii and its adulterant Swertia species by 5S rRNA gene spacer.
Man-Tang YU ; Ka-Lok WONG ; Yu-Ying ZONG ; Pang-Chui SHAW ; Chun-Tao CHE
China Journal of Chinese Materia Medica 2008;33(5):502-504
OBJECTIVEThis research focused on analyzing the differences of 5S rRNA gene spacer sequences on Swertia mussotii and its commonly used adulterants, including S. franchetiana, S. wolfangiana and S. chirayita.
METHODDNA was extracted from the collected Swertia samples. 5S rRNA intergenic spacers were amplified by PCR, sequenced and analyzed.
RESULT5S rRNA gene spacer sequences were different between S. mussotii and its other three adulterants. Sequence divergence among species ranged from 30.6% to 65.0%.
CONCLUSION5S rRNA spacers may be used as molecular authentication markers to differentiate S. mussotii and other commonly used Swertia adulterants. This result provides reliable and simple reference for the authentication of Swertia genus species.
Base Sequence ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Ribosomal, 5S ; genetics ; Sequence Homology, Nucleic Acid ; Swertia ; classification ; genetics
10.Analysis of human papillomavirus type 61, 83 and 84 MY fragments and preparation of standard samples.
Bao-Chen WANG ; Xiao-Li WANG ; Feng-Xia XUE ; Hong-Tu LIU
Chinese Journal of Experimental and Clinical Virology 2010;24(2):107-109
OBJECTIVETo investigate the mutations in nucleotide and amino acid level in HPV61, 83 and 84 Shanxi isolates.
METHODSAmplified fragments of HPV61, 83 and 84 from human papillomavirus (Human papillomavirus, HPV) molecular epidemiologic survey of Shanxi Province using HPV consensus primers MY09/11 were cloned in pMD18-T vector, and the plasmids were sequenced, then nucleotide sequences and amino acid sequences were analyzed.
RESULTSHPV61 and HPV83 isolates were consistent with reference strains U31793 and AF151983 in nucleotide sequences; four mutations of nucleotide (C6760T, T6931C, T6951C and C6987A) were found in HPV84 isolate compared with reference strain AF293960, among which C6987A resulted in D441E and the amplified sensitivity of standard sample of HPV61 using primers MY09/11 was higher than that of HPV83 and 84.
CONCLUSIONHPV61 and HPV83 isolates were consistent with reference strains, four mutations of nucleotide and one mutation of amino acid were found in HPV84,the amplified sensitivity of standard sample of HPV61 using primers MY09/11 was the highest among those three isolates.
Base Sequence ; Genotype ; Humans ; Molecular Sequence Data ; Mutation ; Papillomaviridae ; genetics ; Papillomavirus Infections ; virology ; Sensitivity and Specificity ; Sequence Homology, Nucleic Acid