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SN/T 2531-2010: Determination of Sapovirus in shellfish and water. Conventional RT-PCR and real-time RT-PCR
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivered in | Standard Title (Description) | Status | PDF |
| SN/T 2531-2010 | English | 369 |
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Determination of Sapovirus in shellfish and water. Conventional RT-PCR and real-time RT-PCR
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SN/T 2531-2010
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Basic data | Standard ID | SN/T 2531-2010 (SN/T2531-2010) | | Description (Translated English) | Determination of Sapovirus in shellfish and water. Conventional RT-PCR and real-time RT-PCR | | Sector / Industry | Commodity Inspection Standard (Recommended) | | Classification of Chinese Standard | C53 | | Classification of International Standard | 07.100.30 | | Word Count Estimation | 14,150 | | Date of Issue | 2010-03-02 | | Date of Implementation | 2010-09-16 | | Quoted Standard | SN/T 1193 | | Regulation (derived from) | National Quality Inspection (2010) 98; industry standard filing Notice 2010 No. 5 (No. 125 overall) | | Issuing agency(ies) | General Administration of Customs | | Summary | This standard specifies the shellfish and water samples such as viruses ordinary Sapporo by RT-PCR and real-time fluorescent RT-RCR method. This standard applies to shellfish and water samples Sapporo qualitative detection of such viruses. | SN/T 2531-2010: Determination of Sapovirus in shellfish and water. Conventional RT-PCR and real-time RT-PCR
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Determination of Sapovirus in shellfish and water.Conventional RT-PCR and real-time RT-PCR
Exit inspection and quarantine industry standard book People's Republic of China
Shellfish and water samples sapovirus common detection method
By RT-PCR and real-time RT-PCR.
Issued on. 2010-03-02
2010-09-16 implementation
People's Republic of China
The State Administration of Quality Supervision, Inspection and Quarantine released
Foreword
Appendix A of this standard is a normative appendix, Appendix B is an informative annex.
This standard is proposed and managed by the National Certification and Accreditation Administration Committee.
This standard was drafted. People's Republic of China Shanghai Entry-Exit Inspection and Quarantine Bureau.
The main drafters of this standard. Panliang Wen, Zhong Shan Lu, Deng Tian Ning, Lu Rong, Zhang Shuya, Li Xiang, Liu Yueming, high piano.
This standard is the first release of the entry-exit inspection and quarantine industry standards.
Shellfish and water samples sapovirus common detection method
By RT-PCR and real-time RT-PCR.
1 Scope
This standard specifies the shellfish and water samples sapovirus ordinary by RT-PCR and real-time RT-PCR method.
This standard applies to shellfish and water samples sapovirus qualitative detection.
2 Normative references
The following documents contain provisions which, through reference in this standard and become the standard terms. For dated references, subsequent
Amendments (not including errata content) or revisions do not apply to this standard, however, encourage the parties to the agreement are based on research
Whether the latest versions of these documents. For undated reference documents, the latest versions apply to this standard.
SN/T 1193 genetic testing laboratory technical requirements
3 Terms and Definitions
The following terms and definitions apply to this standard.
3.1
When the number of cycles of each reaction tube fluorescent signal reaches the set value of the field experienced.
3.2
Recombinant molecule comprising a virus-specific cDNA fragments can be detected as a virus PCR positive control.
4 Method summary
Viral RNA was extracted shellfish sample with a suitable lysis buffer (such as Tri-reagent), and according to sapovirus RNA3 'end
Poly (A) structure, connecting with Oligo (dT) 25 magnetic beads specific adsorption sapovirus purified RNA. Water samples of the virus into the
Line after enrichment using a suitable method for extracting and purifying viral RNA. Performed using an ordinary fluorescent RT-PCR or real-time RT-PCR.
Testing. In this study, by constructing a plasmid standard molecules (plasmid molecule contains one copy of each amplified fragment), determine sapovirus ordinary RT-
PCR system for the detection limit of 100 copies, real-time fluorescent RT-PCR system, the detection limit was 10 copies.
5 Reagents
All experimental reagents were of analytical grade; unless otherwise stated, the test water is distilled or deionized water.
5.1 positive samples. sapovirus, -80 ℃ refrigerator; or a plasmid containing sapovirus standard molecular fragment of virus detection, -20 ℃ ice
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5.2 glycine buffer. see Appendix A. 1.1.
5.3 PEG8000 solution. See Appendix A. 1.2.
5.4 lysate. Tri-reagent or other equivalent lysate (eg. TRIzol).
5.5 Oligo (dT) 25 Beads. Dynabeads-oligo (dT) 25 or equivalent products.
5.6 ultrapure water (RNase-free and DNase contamination). See Appendix A. 2.3.
5.7 75% ethanol. See Appendix A. 2.4.
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