Development of a GeXP assay for simultaneous differentiation of the H7 subtype and five NA subtypes of avian influenza viruses
10.3969/j.issn.1002-2694.2024.00.094
- VernacularTitle:同时鉴别诊断H7亚型和5种NA亚型禽流感病毒GeXP检测方法的建立
- Author:
Si-Si LUO
1
;
Zhi-Xun XIE
;
Meng LI
;
Dan LI
;
Li-Ji XIE
;
Sheng WANG
;
Min-Xiu ZHANG
;
Jiao-Ling HUANG
;
Zhi-Qin XIE
;
Ting-Ting ZENG
;
Yan-Fang ZHANG
Author Information
1. 广西壮族自治区兽医研究所/广西兽医生物技术重点实验室/农业农村部中国(广西)—东盟跨境动物疫病防控重点实验室,南宁 530001
- Keywords:
H7 subtype;
NA subtype;
avian influenza virus;
GeXP;
high-throughput
- From:
Chinese Journal of Zoonoses
2024;40(7):670-677
- CountryChina
- Language:Chinese
-
Abstract:
Cases of human infection with H7 subtype avian influenza virus(AIV)combined with five NA subtypes(N2,N3,N4,N7,and N9)have been reported.This study was aimed at establishing a method for simultaneous detection and dif-ferential diagnosis of H7 and five NA subtypes of AIV.Seven pairs of specific primers were designed according to the conserved sequences of the HA gene of H7 subtype AIV,the NA gene of five NA AIV subtypes,and the M gene of all AIV subtypes.A high-throughput GeXP typing method was established for simultaneous detection of the H7 subtype and the five NA subtypes of AIV by using GeXP multiple gene expression and capillary electrophoresis analysis technology.The specificity and sensitivity of the method were determined,and clinical samples were tested.The specificity results indicated that this method was able to simultaneously detect seven target genes in a single tube;each pair of specific primers was able to detect the corresponding AIV subtype,and the universal detection primers were able to detect all subtypes of AIV,with no cross-reaction with other common avian disease pathogens.Sensitivity results demonstrated that this method was able to simultaneously detect seven target genes with a threshold detection limit was 100 copies/μL.The detection results for 150 clinical samples were consistent with those of viral isolation and identification.The high-throughput GeXP method for simultaneous differential diagnosis of the H7 subtype and five subtypes of AIV established in this study has advantages of high specificity,high sensitivity,rapidity,and simplicity,thus providing a new detection method for the effective prevention and control of AIV.
