副结核分枝杆菌重组酶介导扩增-侧向流试纸条检测方法的建立与初步应用

doi:10.11843/j.issn.0366-6964.2024.07.043

Abstract

Abstract:

To establish a rapid and accurate method for detection of Mycobacterium avium subsp. paratuberculosis (MAP), primers and probes were designed according to the specific multicopy insert element IS900 of MAP. A recombinase-aid amplification combined with lateral flow dipstick (RAA-LFD) method was established for visual detection of MAP in the field within 30 min at 35 ℃ after being optimized. The assay was specific with the target sequence of MAP with a detection limit up to 10 fg·μL^-1. One hundred and forty-nine stool samples were tested by this method, showing 100% sensitivity with the PCR. In short, the RAA-LFD assay developed in this study was a rapid, reliable method which can be broadly used.

Key words: Mycobacterium avium subsp. paratuberculosis, recombinase-aid amplification, lateral flow dipstick

CLC Number:

S852.618

Yue NAN, Meizhen LONG, Yuanzhi WANG, Yiduo LIU, Xiangmei ZHOU. Establishment and Preliminary Application of Recombinase-mediated Amplification-sidestream Detection for Detection of Mycobacterium paratuberculosis[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 3255-3260.

Figures/Tables 5

Table 1

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References 17

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引物/探针Primers/Probe	序列(5′ → 3′) Sequences
F	CGCTGCTGGAGTTGATTGCGGCGGTGAC
R	GCGTCTTTGGCGTCGGTCTTGCCTTCGC-Boitin
nfo-Probe	FAM-CGCTGCTGGAGTTGATTGCGGCGG-dSpacer- TGACGACGTTGGCCGATGGAGGCGAGGTCACGTGGGCGATCGACCTCAACGCCGGCGGC GCCGCGTTGCTGATCGCCTTGCTCATCGCTGCCGGGCAGCGGCTGCTTTATATTCCCGG GCGCACGGTCCATCACGCCGCGGGTAGTTACCGCGGCGAAGGCAAGACCGACGCCAAA GACGC-C3-spacer

RAA-LFD	PCR		总样本数 Total number of samples
RAA-LFD	阳性样本数 Number of positive samples	阴性样本数 Number of negative samples	总样本数 Total number of samples
阳性样本数 Number of positive samples	33	22	55
阴性样本数 Number of negative samples	0	94	94
总样本数 Total number of samples	33	116	149

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Establishment and Preliminary Application of Recombinase-mediated Amplification-sidestream Detection for Detection of Mycobacterium paratuberculosis

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