Rapid detection of noroviruses in fecal samples and shellfish by nucleic acid sequence-based amplification
文献类型：期刊论文
| 作者 | Kou, Xiaoxia; Wu, Qingping; Zhang, Jumei; Fan, Hongying |
| 刊名 | Journal of microbiology
 |
| 出版日期 | 2006-08-01 |
| 卷号 | 44期号:4页码:403-408 |
| 关键词 | Detection Noroviruses Nasba Shellfish |
| ISSN号 | 1225-8873 |
| 通讯作者 | Wu, qingping() |
| 英文摘要 | The purpose of this study was to determine the efficacy of a nucleic acid sequence-based amplification (nasba) method of detecting noroviruses in artificially and naturally contaminated shellfish. we used 58 fecal samples that tested positive for noroviruses with electron microscopy (em) to develop an nasba assay for these viruses. oligonucleotide primers targeting the polymerase coding region were used to amplify the viral rna in an isothermal process that resulted in the accumulation of rna amplicons. these amplicons were detected by hybridization with digoxigenin-labeled oligonucleotide probes that were highly specific for genogroup i (gi) and genogroup ii (gii) of noroviruses. the expected band of 327 bp appeared in denaturing agarose gel without any nonspecific band. the specific signal for each amplicon was obtained through northern blotting in many repeats. all fecal samples of which 46 (79.3%) belonged to gii and 12 (20.6%) belonged to gi were positive for noroviruses by em and by nasba. target rna concentrations as low as 5 pg/ml were detected in fecal specimens using nasba. when the assay was applied to artificially contaminated shellfish, the sensitivity to nucleic acid was 100 pg/1.5 g shellfish tissue. the potential use of this assay was also confirmed in naturally contaminated shellfish collected from different ponds in guangzhou city of china, of which 24 (18.76%) out of 128 samples were positive for noroviruses; of these, 19 (79.6 %) belonged to gii and 5 (20.4%) belonged to gi. the nasba assay provided a more rapid and efficient way of detecting noroviruses in fecal samples and demonstrated its potential for detecting noroviruses in food and environmental samples with high specificity and sensitivity. |
| WOS关键词 | HEPATITIS-A VIRUS ; REVERSE TRANSCRIPTION-PCR ; NORWALK-LIKE VIRUSES ; TIME RT-PCR ; UNITED-STATES ; GENOGROUP-I ; ACUTE GASTROENTERITIS ; FOOD-BORNE ; OUTBREAKS ; ASSAY |
| WOS研究方向 | Microbiology |
| WOS类目 | Microbiology |
| 语种 | 英语 |
| WOS记录号 | WOS:000240266700005 |
| 出版者 | MICROBIOLOGY SOC KOREA |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2375308 |
| 专题 | 武汉病毒研究所 |
| 通讯作者 | Wu, Qingping |
| 作者单位 | 1.Guangdong Inst Microbiol, Guangdong Provincial Key Lab Microbial Culture Co, Guangzhou 510070, Peoples R China 2.Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China 3.Grad Univ Chinese Acad Sci, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Kou, Xiaoxia,Wu, Qingping,Zhang, Jumei,et al. Rapid detection of noroviruses in fecal samples and shellfish by nucleic acid sequence-based amplification[J]. Journal of microbiology,2006,44(4):403-408. |
| APA | Kou, Xiaoxia,Wu, Qingping,Zhang, Jumei,&Fan, Hongying.(2006).Rapid detection of noroviruses in fecal samples and shellfish by nucleic acid sequence-based amplification.Journal of microbiology,44(4),403-408. |
| MLA | Kou, Xiaoxia,et al."Rapid detection of noroviruses in fecal samples and shellfish by nucleic acid sequence-based amplification".Journal of microbiology 44.4(2006):403-408. |
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来源：武汉病毒研究所
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