Crops ›› 2021, Vol. 37 ›› Issue (6): 62-66.doi: 10.16035/j.issn.1001-7283.2021.06.010

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Development of a Double Test Strip for the Detection of Potato Virus X and Y

Zhang Wei(), Li Zhixin, Zhao Xue, Zhang Jinpeng, Fu Chunjiang, Yu Qianqian, Liu Weiping   

  1. Keshan Branch of Heilongjiang Academy of Agricultural Sciences, Qiqihar 161600, Heilongjiang, China
  • Received:2020-12-03 Revised:2021-10-18 Online:2021-12-15 Published:2021-12-16

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Abstract:

In order to detect potato virus X (PVX) and Y (PVY) rapidly in potato seed production field, a PVX-PVY test strip was developed by colloidal gold labeling and immunochromatography. Colloidal gold solution was prepared by trisodium citrate reduction method with the maximum absorption value at the wavelength of 526nm and the diameter of gold particles was about 25nm. In the strip test, colloidal gold-labeled polyclonal rabbit antibody against PVX and PVY were sprayed on the glass fiber as the detection antibody, and goat anti-rabbit IgG at the control line and anti-PVX/ anti-PVY at the test line on the nitrocellulose membrane of the test strip were served as the capture antibody. The results showed that the two kinds of viruses could be simultaneously detected within 2min. The detection was still feasible when the PVX sample was diluted by 106(W/V), the PVY sample was diluted by 105 (W/V), which was more sensitive to PVX detection. The test strip showed no cross effect when tested by other common four viruses (PVS, PLRV, PVA, and PVM) samples. DAS-ELISA was used to detect the consistency of potato leaves collected in the field. The strip has the characteristics of simple operation and quick reaction, especially suitable for field and port inspection.

Key words: Potato virus X, Potato virus Y, Double test strip

Fig.1

Colloidal gold absorption spectrum"

Fig.2

Colloidal gold particle morphology under transmission electron microscope (Bar=500nm)"

Fig.3

Specific detection of six common viruses by strips C: control line, X: PVX test line, Y: PVY test line, the same below"

Fig.4

Test strips detect PVX and PVY sap"

Table 1

The results of two methods"

检测方法
Test method		 样本数
Sample number		 PVX PVY
阳性
Positive		 阴性
Negative		 阳性率
Positive rate (%)		 阳性
Positive		 阴性
Negative		 阳性率
Positive rate (%)
双重病毒检测试纸条
Double virus test strip		 93 12 81 12.90 26 67 27.96
DAS-ELISA 93 14 79 15.05 29 64 31.18

Table 2

Comparison of the compatibility between the double virus test strip and DAS-ELISA test for PVX and PVY"

病毒
Virus		 方法
Method		 DAS-ELISA 合计
Total		 符合率
Coincidence rate (%)		 Kappa值
Kappa value
+ -
PVX 双重病毒检测试纸条Double virus test strip + 12 0 12 97.85 0.91
- 2 79 81
合计Total 14 79 93
PVY 双重病毒检测试纸条Double virus test strip + 26 0 26 96.77 0.92
- 3 64 67
合计Total 29 64 93

Fig.5

Stability test results of strip a: negative sample, b: PVX positive sample, c: PVY positive sample, d: PVX and PVY positive sample"

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