Establishment of a dual LFD-RPA rapid test for Mycoplasma hyorhinis and My-coplasma hyopneumoniae in swine
10.16303/j.cnki.1005-4545.2025.11.11
- VernacularTitle:猪鼻支原体和猪肺炎支原体双重LFD-RPA快速检测方法的建立
- Author:
Qi JIA
1
;
Li WANG
;
Hanzhu WANG
;
Jingjing SONG
;
Jing SUN
;
Hui LI
;
Feng LI
;
Kai-shun HAN
;
Zhixin FENG
;
Shuguang LI
Author Information
1. 山东省滨州畜牧兽医研究院,山东滨州 256600
- Publication Type:Journal Article
- Keywords:
Mycoplasma hyorhinis;
Mycoplasma hyopneumoniae;
recombinase polymerase amplifi-cation;
lateral flow dipstick
- From:
Chinese Journal of Veterinary Science
2025;45(11):2380-2386
- CountryChina
- Language:Chinese
-
Abstract:
In order to establish a simple,sensitive and specific diagnostic method for the simultane-ous detection of Mycoplasma hyorhinis(Mhr)and Mycoplasma hyopneumoniae(Mhp)in swine,specific primers and probes were designed using the Mhr p37 and Mhp p36 gene sequences as the target genes,and the dual LFD-RPA rapid test was established by screening the primers and probes,optimizing primer ratios and evaluating its effectiveness through the sensitivity,reproduc-ibility and clinical sample testing.The sensitivity,specificity,reproducibility and clinical samples were evaluated.The results showed that the established dual LFD-RPA assay could complete the amplification in 15 min at 39 ℃,and its optimal primer ratio was 1.6∶0.8,and the lowest detection limits were up to 3.63 and 3.60 copies/μL,respectively;the reproducibility was stable;and there was no cross-reactivity with Pasteurella multocida,Bordetella bronchiseptica,Haemophilus pa-rasuis,Actinobacillus pleuropneumoniae,Escherichia coli.The test successfully established a dual LFD-RPA assay,which can detect Mhr and Mhp simultaneously,and is simple,sensitive and spe-cific without relying on specialized equipment,and is suitable for carrying out on-site rapid diagno-sis of Mhr and Mhp.
