GB/T 14926.8-2026Laboratory animals - Method for the examination of Mycoplasma spp. (English PDF)

Foreword

This document complies with the provisions of GB/T 1.1-2020 "Standardization Work Guidelines Part

1.Structure and Drafting Rules of Standardization Documents". Drafting. This document is Part 8 of GB/T 14926.GB/T 14926 has already published the following parts.

---Detection method for Salmonella in laboratory animals (GB/T 14926.1);

---Detection method for Yersinia in laboratory animals (GB/T 14926.3);

---Detection Method for Pathogenic Fungi in Laboratory Animal Skin (GB/T 14926.4);

---Detection method for Pasteurella multocida in laboratory animals (GB/T 14926.5);

---Detection method for Bordetella bronchiseptica in laboratory animals (GB/T 14926.6);

---Methods for the detection of mycoplasma in laboratory animals (GB/T 14926.8);

---Detection method for Corynebacterium tumefaciens in laboratory animals (GB/T 14926.9);

---Detection method for Tyzer pathogens in laboratory animals (GB/T 14926.10);

---Detection method for Escherichia coli O115a,c.K(B) in laboratory animals (GB/T 14926.11);

---Detection method for Pasteurella pneumophila in laboratory animals (GB/T 14926.12);

---Detection method for Klebsiella pneumoniae in laboratory animals (GB/T 14926.13);

---Detection method for Staphylococcus aureus in laboratory animals (GB/T 14926.14);

1 Scope

GB/T 14926.8-2026 is the Chinese national standard covering detecting Mycoplasma in a laboratory animal colony. Mycoplasma pulmonis produces chronic respiratory disease that quietly changes every physiological measurement made on the animals, which is worse for research than an outbreak that kills them. The method fixes the sampling, the detection technique and its controls, and the interpretation. It replaces GB/T 14926.8-2001, one of the parts of GB/T 14926 revised together in this batch. It was issued on 28 January 2026 and has been in force since 1 May 2026, replacing GB/T 14926.8-2001. The document is under the responsibility of the Ministry of Science and Technology. This page is published from the official record of the 2026 edition; the clause text of a standard this recent is not yet in circulation, and the figures, limits and tables it contains are those of the document itself, delivered in full with the English translation.

This document describes a method for detecting mycoplasma in laboratory animals. This document applies to the detection of mycoplasma in laboratory animals such as mice and rats, as well as in vitro inoculum and environmental samples of laboratory animals.

2 Normative references

The contents of the following documents, through normative references within the text, constitute essential provisions of this document. Dated citations are not included. For references to documents, only the version corresponding to that date applies to this document; for undated references, the latest version (including all amendments) applies. This document.

GB/T 14926.42-2001 Collection of Specimens for Bacteriological Examination of Laboratory Animals

GB/T 14926.43 Laboratory animal bacteriological detection staining methods, culture media and reagents

GB/T 14926.50 Enzyme-linked immunosorbent assay (ELISA) for laboratory animals Pharmacopoeia of the People's Republic of China

3 Terms and Definitions

This document does not contain any terms or definitions that need to be defined.

4.Principles Mycoplasma can form distinctive colonies on culture media, which show a characteristic blue color after Dienes staining; this can be verified using an enzyme-linked immunosorbent assay (ELISA). When detecting serum antibodies using ELISA, the antigen-antibody-enzyme-labeled complex catalyzes a colorimetric reaction, and the color intensity is directly proportional to the antibody concentration; targeting specific antibodies... Primers and probes were designed based on the conserved genes of the original organism, and the presence of nucleic acids was determined by specifically amplifying DNA fragments using polymerase chain reaction (PCR).

5 Culture media and reagents

5.1 Mycoplasma semi-fluid culture medium.

5.2 Mycoplasma liquid culture medium.

5.3 Mycoplasma solid culture medium.

5.4 Dienes staining solution.

5.5 The ELISA antigen was prepared according to the following procedure.

b) Centrifuge the obviously turbid culture medium at 8000 r/min for 30 min, and wash the precipitate with phosphate-buffered saline (PBS) under the same conditions. 3 times;

c) The above precipitate was resuspended in PBS, and after being broken up by sonication, the supernatant was used as the ELISA antigen.