GB/T 14926.62-2026 Laboratory animal—Method for examination of simian immunodeficiency virus (SIV) English, Anglais, Englisch, Inglés, えいご
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ICS 13.220.10
CCS H 57
National Standard of the People's Republic of China
GB/T 14926.62-2026
Replaces GB/T 14926.62-2001
Laboratory animal - Method for examination of simian immunodeficiency virus (SIV)
实验动物 猴免疫缺陷病毒检测方法
Issue date: 2026-01-28 Implementation date: 2027-02-01
Issued by the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China
the Standardization Administration of the People's Republic of China
Contents
Foreword
Introduction
1 Scope
2 Normative References
3 Terms and Definitions
4 Principle
5 Reagents and Equipment
6 Test Methods
7 Result Interpretation
8 Result Reporting
Laboratory Animals — Detection Method for Simian Immunodeficiency Virus
1 Scope
This document describes detection methods for Simian immunodeficiency virus (SIV).
This document applies to the detection of SIV antibodies and nucleic acid in experimentally housed monkeys.
2 Normative References
The following documents are referred to in the text in such a way that some or all of their content constitutes requirements of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.
GB/T 14926.50 Laboratory animal – Enzyme-linked immunosorbent assay
GB/T 14926.52 Laboratory animal – Immunofluorescence assay
3 Terms and Definitions
No terms and definitions are required for this document.
4 Principle
4.1 Principle of Antibody Detection
Based on immunological principles, SIV antigen is used to detect SIV antibodies in monkey serum.
4.2 Principle of Nucleic Acid Detection
Specific primers and probes are designed and synthesized based on the conserved SIV gag gene sequence of Simian immunodeficiency virus to amplify target fragments. Real-time quantitative reverse transcription polymerase chain reaction (real-time quantitative RT-PCR) is used to detect whether viral nucleic acid components are present in the sample.
5 Reagents and Equipment
5.1 Reagents
5.1.1 Enzyme-linked Immunosorbent Assay (ELISA) Antigen
5.1.1.1 Specific Antigen
Infect CEM-174 cells with SIV. Harvest the culture medium when significant cell fusion is observed. Freeze-thaw three times or sonicate. Remove cell debris by low-speed centrifugation. Concentrate the supernatant by ultracentrifugation to prepare the ELISA antigen.
Alternatively, SIV-specific recombinant protein antigen produced by genetic engineering.
5.1.1.2 Normal Antigen
Supernatant obtained by freeze-thawing and disrupting uninfected CEM-174 cells, followed by low-speed centrifugation to remove cell debris.
5.1.2 Antigen Slides
Infect CEM-174 cells with SIV. When cytopathic effect (CPE) reaches ++ to +++, centrifuge at 800 r/min for 5 min to 10 min. Wash with PBS and prepare smears. Air dry at room temperature while simultaneously irradiating with UV light at a distance of 20 cm for 30 minutes. Fix with cold acetone for 10 minutes. Store at -20°C or below.
5.1.3 Positive Serum
Monkey serum immunized with SIV antigen or serum from monkeys infected with SIV virus.
5.1.4 Negative Serum
Normal monkey serum free from SIV infection.
5.1.5 Enzyme Conjugate
Horseradish peroxidase-labeled goat or rabbit anti-monkey IgG antibody.
5.1.6 Fluorescent Conjugate
Fluorescein isothiocyanate-labeled goat or rabbit anti-monkey IgG antibody.
5.1.7 Nucleic Acid Detection Reagents
5.1.7.1 DEPC-treated RNase-free water or commercially available RNase-free water.
5.1.7.2 RNA extraction reagent: commercially available reagent.
5.1.7.3 Reverse transcription reagent: commercially available reagent.
5.1.7.4 Real-time quantitative PCR master mix: commercially available reagent.
5.1.7.5 Positive control: Tissue sample or cell culture containing SIV virus nucleic acid, or a plasmid containing the SIV-specific sequence.
5.1.7.6 Negative control: Tissue sample or cell culture not containing SIV virus nucleic acid.
5.1.7.7 Primers and probes: Synthesize according to the sequences in Table 1. Prepare primers and probes as 10 μmol/L stock solutions. Store at -20°C.
5.2 Equipment
5.2.1 Microplate reader.
5.2.2 Fluorescence microscope.
5.2.3 37°C incubator or water bath.
5.2.4 Real-time quantitative PCR instrument, PCR instrument.
5.2.5 Centrifuge.
5.2.6 Biosafety cabinet.
5.2.7 Clean bench (laminar flow hood).
5.2.8 Other equipment: Micropipettes, centrifuge tubes, polystyrene microplates, PCR tubes or plates, etc.
6 Test Methods
6.1 Use the ELISA method for serological detection, operating according to GB/T 14926.50.
6.2 Use the indirect immunofluorescence assay (IFA) method for serological detection, operating according to GB/T 14926.52.
6.3 Use the real-time quantitative RT-PCR method for SIV nucleic acid detection, operating according to Appendix A.
7 Result Interpretation
7.1 Interpretation of Antibody Detection Results
