SN/T 5105-2019 in English

VALID

Detection method of Rotavirus( group B ) by real-time fluorescenceRT-PCR at frontier ports

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

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Standard No:	SN/T 5105-2019
Document status:	VALID
Title in English:	Detection method of Rotavirus( group B ) by real-time fluorescenceRT-PCR at frontier ports
Title in Chinese:	国境口岸轮状病毒（B组）实时荧光RT-PCR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:	detection method traditional rt-pcr real-time fluorescence rt-pcr detection time detection sensitivity open-cap detection closed-tube detection
Related Topics:	Two rounds of rt-pcr pcr port real time Fluorescence detection Fluorescence detection Fluorescence curve fluorescent signal Fluorescent real-time PCR pcr detection real-time fluorescence Real-time fluorescent PCR detection method for Borrelia burgdorferi at border ports PCR 40 rounds Isothermal fluorescent PCR pcr sample rt detection X fluorescence detection at home and abroad What does rt-pcr detect rt-pcr several per group border port relative fluorescence pcr Fluorescence curve border port pcr detection fluorescence response Multiplex PCR real-time fluorescence quantitative PCR Rift Valley Fever Virus RT-PCR and Real-time Fluorescent RT-PCR Detection Methods fluorescent signal rt detection Surveillance regulations for rotavirus infection at ports of entry and exit virus fluorescence method Fluorescence detection method by flow cytometry fluorescent pcr peak rt pcr Multiplex fluorescent PCR Real-time fluorescent PCR Column fluorescence detection method round mouth real-time fluorescence detection When detecting fluorescence spectra? pcr rna ????pcr?? Q7 fluorescent PCR Domestic fluorescence quantitative PCR Rotavirus group rt pcr gene pcr standard When histamine is detected q6 fluorescent pcr amplified pcr Toxic detection at any time RT-PCR method to detect non- fluorescent fluorescent probe Fluorescence data Domestic water quality virus detection virus group Rotavirus real-time fluorescence real-time pcr analysis method real-time fluorescent quantitative pcr and pcr Fluorescent real-time PCR Q5 fluorescent PCR Phenomenal fluorescence How to detect virus by fluorescent quantitative PCR Fluorescence PCR instrument usage method rt in pcr Methods for detecting fluorescence Tissue virus PCR detection flow real-time fluorescence Fluorescence Signal Standard plate time fluorescence rt/pcr domestic pcr pcr detection method report fluorescent histamine Fluorescence detection by pcr fluorescent pcr q6 real-time fluorescence Photoelectric detection abroad pcr flow cytometry fluorescence detection fluorescent pcr iso Poison b real-time fluorescent quantitative pcr and pcr fluorescence real-time Virus + five fluorescent Isothermal PCR Real-time fluorescent PCR Fluorescence PCR detection series Fluorophore Fluorescence quenching France RT 2012 U.S. drug testing methods Histamine fluorescence detection method Fluorescein detection method Spectral disease detection method Fluorescent group national standard Fluorescence and other detection methods Fluorescence detector method

Introduction

Analysis of the core content of the standard

This standard uses real-time fluorescence RT-PCR technology to detect rotavirus group B specific gene fragments, with a detection sensitivity of 10³ copies/μL, which is 100 times higher than that of traditional methods. The key technical features include:

Technical parameters	Traditional RT-PCR	Real-time fluorescence RT-PCR
Detection time	4-6 hours	2 hours
Sensitivity	10⁵ copies/μL	10³ copies/μL
Contamination risk	High (open-cap detection required)	Low (closed-tube detection)

Key technical principles

The molecular beacon probe technology (sequence: 5'-FAM-CGATCG...GATCG-DABCYL-3') is used, and its stem-loop structure design makes:

Free state: the distance between the fluorescent group and the quenching group is <10Å, and the fluorescence quenching efficiency is >99%
Target binding state: the probe unfolds to increase the distance between FAM and DABCYL to >30Å, and the fluorescence recovery rate is >80%

Detection process specifications

Typical operation cases

The detection example of Zhuhai Port in 2020 shows:

Sample pretreatment uses the Trizol method to extract RNA, and the recovery rate is ≥85%
The reaction system contains 2×BufferⅢ 12.5μL, 1μL of each primer (10μmol/L)
Cycling parameters: 42℃ 5min reverse transcription→95℃ 10s pre-denaturation→40 PCR cycles

Result judgment criteria

Ct value range	Judgment result	Handling suggestions
≤35	Positive	Directly report detection
35-40	Suspected positive	Need to concentrate and retest
No amplification	Negative	Combined with clinical judgment

Standard implementation recommendations

Quality control requirements: Each batch of testing must include a positive control (Ct≤30) and a blank control
Equipment calibration: Fluorescence quantitative PCR instrument requires optical path calibration every quarter
Staff training: Operators should pass the biosafety level 3 protection assessment

More samples: NB/T 47013.2-2015 JC/T 2453-2018 GB 18306-2015

Sample only — not a preview of SN/T 5105-2019

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