SN/T 2120-2014Quarantine protocol for epizootic ulcerative syndrome (English PDF)

Foreword

This standard was drafted in accordance with the rules given in GB/T 1.1-2009.

This standard replaces SN/T 2120-2008 "Technical Specification for Quarantine of Epidemic Ulcers".

Compared with SN/T 2120-2008, the main technical changes of this standard are as follows.

--- Increased sampling;

--- Increased clinical symptoms;

--- Modified the induction of secondary migration spores;

--- Modified the method of processing samples in PCR;

--- Modified the PCR test primers;

--- Adjusted and optimized the PCR test conditions;

--- Modified the result judgment.

This standard is proposed and managed by the National Certification and Accreditation Administration.

This standard was drafted. Shenzhen Entry-Exit Inspection and Quarantine Bureau of the People's Republic of China, Shenzhen Institute of Inspection and Quarantine, Chinese People Guangdong Entry-Exit Inspection and Quarantine Bureau.

1 Scope

This standard specifies the pathogen isolation, identification and diagnosis methods for various freshwater and brackish water fish epidemic ulcer syndromes.

This standard applies to the quarantine and disease surveillance of various freshwater and brackish water fish epidemic ulcer syndrome.

2 Normative references

The following documents are indispensable for the application of this document. For dated references, only dated versions apply to this article.

Pieces. For undated references, the latest edition (including all amendments) applies to this document.

GB/T 6682 Analytical laboratory water specifications and test methods GB/T 18088 entry and exit animal quarantine sampling GB 19489 General requirements for laboratory biosafety

3 Overview

Epizootic Ulcerative Syndrome, also known as red spot disease (RSD), fungal granuloma (MG), mildew Bacterial ulcer (UM), a wild and cultured freshwater and half caused by Aphanomyces invadans or A.piscicida the seasonal epidemics of saltwater fish, mainly characterized by invasive filaria infection and necrotic ulcer disease, characteristically causing granulomatous should be referred to as EUS (see Appendix A).

4 Reagents and materials

4.1 Water. Water used for PCR (including nucleic acid extraction) is treated with DEPC to remove DNase and RNase. The remaining water should meet GB/T 6682 grade 1 water specification.

4.2 Cryptosporidium reference strain. provided by the designated unit.

4.3 Taq DNA polymerase. Store at -20 °C, do not freeze or thaw repeatedly or drastic changes in temperature.

4.4 dNTPs. containing 10 mmol/L of dCTP, dGTP, dATP and dTTP.

4.5 Primers. Two pairs of primers with a concentration of 40 µmol/L. The sequence is.

Ainvad-2F. 5'-TCA-TTG-TGA-GTG-AAA-CGG-TG-3' Ainvad-ITSR1. 5'-GGC-TAA-GGT-TTC-AGT-ATG-TAG-3' A 234 bp DNA fragment in the ITS1rDNA gene of the genus Aspergillus was amplified.

ITS11. 5'-GCC-GAA-GTT-TCG-CAA-GAA-AC-3' ITS23. 5'-CGT-ATA-GAC-ACA-AGC-ACA-CCA-3' A 550 bp DNA fragment in the ITS1rDNA gene of the genus Aspergillus was amplified.

4.6 Anhydrous ethanol. Analytically pure, pre-cooled to -20 °C before use.

4.7 10% formalin.

4.8 Alcohol. 70%, 80%, 95% and anhydrous alcohol.