基于酶促重组酶扩增的非洲猪瘟病毒检测方法

doi:10.11843/j.issn.0366-6964.2024.09.046

Abstract

Abstract:

This experiment was conducted to establish a rapid nucleic acid detection method for African swine fever virus (ASFV) based on enzymatic recombinase amplification (ERA). We designed ERA probes and primers specific for the conserved sequence of ASFV B646L gene, and optimized the reaction conditions in order to establish an ERA method for the detection of ASFV DNA under isothermal conditions. The ERA method for ASFV demonstrated high specificity and no cross-reaction with other pathogens; the CV was less than 10%, indicative of good reproducibility. The lowest detection limit for the ERA method is 85 copies·μL^-1; comparison with the World Organization for Animal Health (WOAH) recommended qPCR diagnostic method for African swine fever (ASF) demonstrated a Kappa value of 0.961, suggestive of high identity with African swine fever qPCR diagnostic method. ERA-based method for the detection of ASFV can be used for the rapid detection of ASFV in the field.

Key words: African swine fever virus, ERA, Nucleic acid detection, Isothermal amplification

CLC Number:

S852.659.1

Lu FENG, Hong TIAN, Haixue ZHENG, Zhengwang SHI, Juncong LUO, Xiaoyang ZHANG, Juanjuan WEI, Jing ZHOU, Huancheng LIAO, Wanying WANG. A Detection Method of African Swine Fever Virus based on Enzymatic Recombinase Amplification[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 4226-4231.

Figures/Tables 4

Fig. 1

Table 1

Fig. 2

Table 2

References 13

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样品编号 Sample No.	Ct值 Ct value				变异系数/% Coefficient of variation
样品编号 Sample No.	重复1 Repetition 1	重复2 Repetition 2	重复3 Repetition 3	标准偏差 Standard deviation	变异系数/% Coefficient of variation
1	9.47	9.41	8.36	0.62	6.87
2	5.57	6.1	5.50	0.33	5.73
3	5.22	4.98	4.53	0.35	7.13
4	6.11	6.2	5.24	0.53	9.01
5	7.69	7.78	8.58	0.49	6.11
6	6.1	6.68	6.24	0.30	4.77
7	9.55	9.81	9.18	0.32	3.33
8	9.89	11.06	12.00	1.05	9.62
9	15.03	16.73	15.84	0.85	5.36
10	11.83	12.04	11.10	0.49	4.23

方法 Method		qPCR
方法 Method		阳性 Positive	阴性 Negative	合计 Total
ERA	阳性Positive	19	0	19
	阴性Negative	1	40	41
	合计Total	20	40	60
Kappa值 Kappa value		0.961

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A Detection Method of African Swine Fever Virus based on Enzymatic Recombinase Amplification

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