GB/T 14926.23-2026 Laboratory animal—Method for examination of Sendai virus（SV） 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 14926.23-2026
Replaces GB/T 14926.23-2001
Laboratory animal - Method for examination of Sendai virus（SV）
实验动物 仙台病毒检测方法
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 Main Equipment
6 Reagents
7 Detection Methods
8 Result Interpretation
9 Result Reporting
Laboratory Animals — Detection Method for Sendai Virus
1 Scope
This document describes detection methods for Sendai virus (SV).
This document applies to the detection of Sendai virus in laboratory animals such as mice, rats, guinea pigs, hamsters, and rabbits.
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 14926.42-2001 Laboratory animal – Bacteriological examination – Collection of specimens
GB/T 14926.50 Laboratory animal – Enzyme-linked immunosorbent assay
GB/T 14926.51 Laboratory animal – Immunoenzyme assay
GB/T 14926.52 Laboratory animal – Immunofluorescence assay
GB/T 14926.54 Laboratory animal – Haemagglutination inhibition test
3 Terms and Definitions
No terms and definitions are required for this document.
4 Principle
4.1 Antibody Detection
Based on immunological principles, SV antigen is used to detect Sendai virus antibodies in the serum of mice, rats, guinea pigs, hamsters, and rabbits. Alternatively, based on the principle that the ability of SV to agglutinate chicken or guinea pig erythrocytes under certain conditions can be inhibited by specific antibodies, SV antibodies in samples can be detected.
4.2 Nucleic Acid Detection
The unique genomic nucleic acid sequences of Sendai virus can be identified by nucleic acid amplification techniques such as polymerase chain reaction (PCR).
5 Main Equipment
5.1 Microplate reader.
5.2 Fluorescence microscope, magnification range 100x to 1000x.
5.3 Ordinary light microscope, magnification range 100x to 1000x.
5.4 37°C incubator or water bath, temperature control range 25°C to 100°C.
5.5 Real-time quantitative polymerase chain reaction (PCR) instrument, temperature accuracy ±0.5°C, temperature uniformity ≤ 1°C, average heating and cooling rate ≥ 1.5°C/s.
6 Reagents
6.1 Enzyme-linked Immunosorbent Assay (ELISA) Antigen
6.1.1 Specific Antigen
Infect the allantoic cavity of 9-day-old specific pathogen free (SPF) chicken embryos with SV. Incubate at 36°C. After 72 hours, chill at 4°C. The next day, aseptically collect the allantoic fluid. Centrifuge at 2000 r/min for 10 min at 4°C. Verify virus specificity and hemagglutination (HA) titer using 0.5% chicken or guinea pig erythrocytes and SV positive serum in HA and hemagglutination inhibition (HI) tests. Concentrate the supernatant by ultracentrifugation to prepare the ELISA antigen.
6.1.2 Normal Antigen
Allantoic fluid from 9-day-old SPF chicken embryos.
6.2 Antigen Slides
Infect baby hamster kidney cells (BHK21) with SV. 2 to 3 days post-infection, when cytopathic effect (CPE) reaches ++ to +++, disperse the cells with trypsin, wash with phosphate-buffered saline (PBS), and prepare smears. Air dry at room temperature. Fix with cold acetone for 10 minutes. Store at -20°C.
6.3 Hemagglutinin
Refer to the preparation of ELISA specific antigen.
6.4 Positive Serum
Antiserum obtained by immunizing SPF mice, rats, guinea pigs, hamsters, or conventional rabbits with SV antigen.
6.5 Negative Serum
Serum from SPF mice, rats, guinea pigs, hamsters, and rabbit serum free from Sendai virus infection.
6.6 Enzyme Conjugate
Horseradish peroxidase, etc. labeled goat or rabbit anti-mouse, rat, guinea pig, hamster IgG antibodies, used for detecting corresponding animal serum antibodies; the aforementioned enzyme-labeled goat anti-rabbit IgG antibody, used for detecting rabbit serum antibodies; the aforementioned enzyme-labeled Staphylococcal Protein A (SPA) can be used for detecting serum antibodies in mice, guinea pigs, hamsters, and rabbits.
6.7 Fluorescent Conjugate
Fluorescein isothiocyanate, etc. labeled goat or rabbit anti-mouse, rat, guinea pig, hamster IgG antibodies, used for detecting corresponding animal serum antibodies; use the aforementioned labeled goat anti-rabbit immunoglobulin G (IgG) antibody for detecting rabbit serum antibodies.
6.8 Real-time Quantitative Reverse Transcription-Polymerase Chain Reaction (RT-PCR) Reagents
6.8.1 Nuclease-free deionized water.
6.8.2 Ribonucleic acid (RNA) extraction reagents or kit.
6.8.3 One-step reverse transcription-real-time quantitative PCR kit or equivalent kit.
6.8.4 Primers and probes: Synthesize primers and probes according to the sequences in
Table 1. Prepare primers and probes as 20 μmol/L stock solutions using ribonuclease (RNase)-free deionized water. Store at -20°C.
7 Detection Methods
7.1 Collect serum according to the serum sampling requirements in 4.3 of GB/T 14926.42-2001. Collect other samples according to Appendix A.
7.2 Use the ELISA method, operating according to GB/T 14926.50.
7.3 Use the immunofluorescence assay (IFA), operating according to GB/T 14926.52.
7.4 Use the immunoenzyme staining assay (IEA), operating according to GB/T 14926.51.
