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

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"

[1] 郭兴启, 冯炘, 李向东, 等. PVY/PVX协生作用对病毒浓度及寄主细胞超微结构的影响. 中国农业科学, 2003(3):281-286,355-356.
[2] 吴兴泉, 张慧聪, 时妍, 等. 我国部分马铃薯产区主要病毒病发生情况调查. 河南农业科学, 2013, 42(7):84-87.
[3] 范国权, 白艳菊, 高艳玲, 等. 我国马铃薯主产区病毒病发生情况调查. 黑龙江农业科学, 2014(3):68-72,87.
[4] 周淑芹, 朱光新. 电子显微镜技术在鉴定筛选马铃薯无毒核心材料中的应用. 黑龙江农业科学, 1996(6):17-20.
[5] 郑世玲, 刘作易. 贵州3种马铃薯病毒的DAS-ELISA检测与分析. 贵州农业科学, 2006(6):42-44.
[6] 罗文彬, 李华伟, 汤浩, 等. 马铃薯5种病毒多重PCR检测技术的建立及应用. 园艺学报, 2015, 42(2):280-288.
[7] 张京宣, 曹秀芬, 宋涛, 等. 植物病毒检测技术研究进展分析. 北京农业, 2011(30):79-80.
[8] Suwussa B, Chayachon A, Warangkana C, et al. Rapid and sensitive lateral flow immunoassay for influenza antigen using fluorescently-doped silica nanoparticles. Microchimica Acta, 2014, 181(1/2):223-230.
doi: 10.1007/s00604-013-1106-4
[9] Li L Y, Jian Z D, Li Y W, et al. Development of a rapid dipstick with latex immunochromatographic assay (DLIA) for diagnosis of schistosomiasis japonica. Parasites and Vectors, 2011, 4:157.
doi: 10.1186/1756-3305-4-157 pmid: 21824443
[10] 纪玲玲, 成巨龙, 朱信宁, 等. 烟草脉带花叶病毒试纸条的制作及田间病株的快速检测. 西北农业学报, 2009, 18(1):180-183.
[11] 赵肖, 白昀, 陈蓉, 等. 猪肺炎支原体抗体胶体金免疫层析检测方法的建立. 中国兽医学报, 2018, 38(9):1693-1698.
[12] 冯忠华, 李跃龙, 陈运勤, 等. 胶体金免疫层析技术及其在食品安全快速检测中的应用. 广东饲料, 2017, 26(12):39-40.
[13] 魏梅生, 刘洪义, 李桂芬, 等. 马铃薯X病毒和马铃薯Y病毒胶体金免疫层析试纸条的研制. 植物保护, 2006(6):139-141.
[14] 陈阳婷, 桑有顺, 冯焱, 等. 双重RT-PCR法快速检测多种马铃薯病毒的研究. 西南农业学报, 2012, 25(1):179-182.
[15] 张微, 李志新, 付春江, 等. 马铃薯S病毒胶体金免疫层析试纸条的研制. 生物技术通报, 2019, 35(12):184-188.
doi: 10.13560/j.cnki.biotech.bull.1985.2019-0287
[16] Kay S. Maize chlorotic mottle machlomovirus and wheat streak mosaic rymovirus concentrations increase in the synergistic disease corn lethal necrosis. Virology, 1998, 242(1):28-38.
doi: 10.1006/viro.1997.8989
[17] 宋西娇, 谢礼, 宣裕吉, 等. 复合侵染的马铃薯花叶病病原诊断. 浙江农业学报, 2018, 30(1):99-105.
[18] 阮小蕾, 邓海滨, 王晓宾, 等. 多重烟草病毒胶体金检测试纸条的研制及应用. 烟草科技, 2018, 51(10):33-38.
[19] 董雅琴, 刘爽, 郑辉, 等. 三种伪狂犬病病毒gE抗体ELISA检测试剂盒的比较. 中国动物检疫, 2017, 34(11):79-81,88.
[20] 陈祺, 黄辉, 唐长玖, 等. 两种不同凝血功能检测方法的相关性和一致性研究. 中国实验血液学杂志, 2017, 25(6):1820-1824.
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