SN/T 5197-2019 in English

VALID

Quarantine protocol for real time PCR of lumpy skin disease virus

Issued on:2019-12-27
Implemented on:2020-07-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $130.00
$127.00

Standard No:	SN/T 5197-2019
Document status:	VALID
Title in English:	Quarantine protocol for real time PCR of lumpy skin disease virus
Title in Chinese:	牛结节疹病毒荧光定量PCR操作规程
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2019-12-27
Implemented on:	2020-07-01
Professional Classification:	SN-Import&Export Inspection
Related Keywords:	lumpy skin disease virus introduction technical background real time pcr traditional pcr standard nodular herpes virus quarantine protocol
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Introduction

Technical background and significance of the standard

As my country's first molecular detection specification for bovine nodular herpes virus (LSDV), this standard fills the technical gap in the rapid detection of poxviral pathogens. When it was released in 2019, LSDV was out of control in the Middle East and Eastern Europe. The implementation of the standard provided key technical support for port epidemic prevention and control.

Analysis of core detection principles

Technical elements	Traditional PCR	Standard qPCR
Detection target	ITR conserved region	ITR specific fragment
Probe design	None	FAM-MGB dual-labeled probe
Sensitivity	10³ copies	10¹ copies

Typical case: In 2020, Chongqing Customs adopted this standard in the quarantine of imported live cattle, and LSDV was detected from a lymph node sample with a Ct value of 32.5. Sequencing verification showed that the homology with the Neethling standard strain was 99.7%.

Key Operation Points

Nucleic Acid Extraction: A validated commercial kit must be used, and tissue samples must be triple frozen-thawed.
Reaction System: The final concentration of the probe is strictly controlled at 0.2μmol/L to avoid quenching effect.
Cycling Parameters: Annealing/extension uses a two-step optimization scheme of 60℃ 34s.

Quality Control System

Standardized establishment of three-level quality control:
1) Equipment calibration: The fluorescence quantitative PCR instrument is required to perform optical module calibration every year.
2) Control setting: Including positive control (Ct<28), negative control and blank control.
3) Critical value verification: 35<Ct≤40 samples must be retested

Implementation and application recommendations

Laboratories should regularly use the Neethling standard strain to verify the method
It is recommended to conduct parallel testing with the OIE recommended method for 3 months as a method transition period
Tissue samples should preferably be selected from high-load organs such as skin nodules and spleen

More samples: JGJ 94-2008 JC/T 2453-2018 GB 50021-2001(2009)

Sample only — not a preview of SN/T 5197-2019

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