SN/T 5125-2019 in English

VALID

Duplex real-time PCR for Vibrio Parahaemol yticus and Vibrio Cholerae in Aquatic aminals

Issued on:2019-09-03
Implemented on:2020-03-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $70.00
$68.00

Standard No:	SN/T 5125-2019
Document status:	VALID
Title in English:	Duplex real-time PCR for Vibrio Parahaemol yticus and Vibrio Cholerae in Aquatic aminals
Title in Chinese:	水生动物副溶血弧菌和霍乱弧菌双重实时荧光检测方法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2019-09-03
Implemented on:	2020-03-01
Professional Classification:	SN-Import&Export Inspection
Related Keywords:	vibrio cholerae vibrio cholerae ompw cy5 vibrio parahaemolyticus vibrio parahaemol yticus vibrio parahaemolyticus toxr fam
Related Topics:	Fluorescence real-time quantitative multiplex fluorescence fluorescent blood Fluorescence detection method Fluorescence detection method pcr vibrio fluorescens real-time fluorescence detection animal blood testing The fluorescence spectrum is messy Fluorescent detection of serum double serum test Bdellovibrio Vibrio parahaemolyticus Vibrio haemolyticus Vibrio haemolyticus dual fluorescence detector vibrio parahaemolyticus pcr Vibrio parahaemolyticus PCR detection Real-time detection of microorganisms in water vibrio parahaemolyticus pcr Vibrio parahaemolyticus detection pcr light detection method In vivo detection of fluorescent animals vibrio parahaemolyticus pcr Vibrio parahaemolyticus PCR detection dual fluorescent receptor Cholera testing room Illegal detection Biochemical identification of Vibrio vulnificus and Vibrio parahaemolyticus Effect of lysozyme on Bacillus parahaemolyticus Bacterial heme detection method Blood heavy metal detection method Vibrio cholerae detection standards Vibrio cholerae detection method Luciferase method for detecting bacteria Animal blood oxygen measurement method Vibrio parahaemolyticus detection method Solution sterility test method plate Animal blood lipid detection method Sterile animal testing methods Hydrogen peroxide detection method Water soluble matter detection method Sterility testing method for finished blood bags Mycelium dry weight measurement method Protoplast dual fluorescence method National testing standard food for Vibrio cholerae Vibrio harveyi testing method Water soluble matter detection Methods for preventing hemolysis using animal serum Testing methods for sterile water Method for detecting Vibrio cholerae suspension How to detect hydrogen peroxide Streptococcus parahaemolyticus culture method Extravascular hemolysis detection method Vibrio parahaemolyticus sampling requirements Vibrio parahaemolyticus testing and storage requirements

Introduction

Analysis of Standard Technical Framework

Detection Object	Target Gene	Probe Fluorescent Labeling	Product Length (bp)
Vibrio cholerae	OmpW	CY5	95
Vibrio parahaemolyticus	ToxR	FAM	98

Core Detection Process

Sample pretreatment: 3% alkaline peptone water for bacterial enrichment (GB 4789.7)
DNA extraction: 10,000r/min centrifugation combined with proteinase K digestion
Reaction system: 2mmol/L MgCl₂, 0.2mmol/L dNTPs
Amplification procedure: 95℃ 1min initial denaturation, 40 cycles (95℃ 15s→60℃ 34s)

Result judgment criteria

When CT value ≤30.0, it is judged as positive:
- FAM channel positive: Vibrio parahaemolyticus is detected
- CY5 channel positive: Vibrio cholerae detected

CT values between 30.0-35.0 need to be retested, and those still in this range are considered suspicious

Technology evolution analysis

Compared with the traditional culture method, this standard:
1. The detection cycle is shortened from 5-7 days to 4 hours
2. The sensitivity is increased to 10² CFU/mL
3. Simultaneous identification of two pathogens is achieved

Implementation recommendations

The laboratory needs to be equipped with ABI 7500 and above fluorescent PCR instruments
It is recommended to set up negative controls (sterile PBS) and positive controls (ATCC standard strains) for each batch
The enrichment time of environmental samples needs to be extended to 18-24 hours

More samples: NB/T 47013.2-2015 YD/T 3934-2021 JJG 21-2008

Sample only — not a preview of SN/T 5125-2019

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