Studies on Techniques of Detecting Papaya Ringspot Virus

CHEN Zhi-Nan; ZHENG Guan-Biao; LIN Kong-Xun; FAN Huai-Zhong; ZHANG Chun-Xia; TIAN Bo

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April 1993

CHEN Zhi-Nan, ZHENG Guan-Biao, LIN Kong-Xun, FAN Huai-Zhong, ZHANG Chun-Xia and TIAN Bo. Studies on Techniques of Detecting Papaya Ringspot Virus[J]. Virologica Sinica, 1993, 8(2).

Citation: CHEN Zhi-Nan, ZHENG Guan-Biao, LIN Kong-Xun, FAN Huai-Zhong, ZHANG Chun-Xia, TIAN Bo. Studies on Techniques of Detecting Papaya Ringspot Virus .VIROLOGICA SINICA, 1993, 8(2) : 164.

番木瓜环斑病毒检测技术的研究

1.
华南农业大学植保系
2.
中国科学院微生物所
3.
中国科学院微生物所广州 510642 深圳动植物检疫局深圳

摘要

本研究在37℃条件下,以硝酸纤维素滤膜(NCM)为载体:3％酪蛋白为封闭剂、辣根过氧化物酶标记的A蛋白为酶标结合物(SPA-HRP)和以稀释100倍兔抗PRV-Ys的IgG为其工作浓度,成功地建立了检测PRV的间接Dot-ELISA。其检测灵敏度达到了较满意的ng水平。同时,本研究以PRV-Ys株系RNA作模板、oligo(dT)~(12-18)作引物、pUC19作cDNA克隆载体和以E.coli JM107作受体,采用缺口翻译(Nick Translation)方法成功地制备了pYs6 cDNA分子探针(插入片段约300bp)。其检测PRV-RNA的灵敏度达到了较理想的pg水平。
- 华南农业大学植保系
- , 中国科学院微生物所
- , 中国科学院微生物所广州 510642 深圳动植物检疫局深圳

Studies on Techniques of Detecting Papaya Ringspot Virus

1.
Plant Protection Department
2.
South China Agriculture University Guangzhou
3.
0642Institute of Microbiol.

Abstract

A sensitive indirect dot-ELISA was developed by using nitrocellulose membrance (NCM) instead of microplate, 3% casin as a blocking agent, horse-radish peroxidase conjugated protein A as an enzyme-labelled complex and a dilution by 100 folds of IgG from antiserum of rabbit to PRV-Ys strain as the working concentration. and was found to b workable for assessing an amount of PRV particles at ng level (ave.10ng/spot, at 37℃). A PRV-RNA cDNA molecular probe (the insert ca. 300bp) was successfully constructed wit...
- Papaya ringspot virus Indirect dot ELISA cDNA molecular probe

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Studies on Techniques of Detecting Papaya Ringspot Virus

1. Plant Protection Department
2. South China Agriculture University Guangzhou
3. 0642Institute of Microbiol.

Abstract: A sensitive indirect dot-ELISA was developed by using nitrocellulose membrance (NCM) instead of microplate, 3% casin as a blocking agent, horse-radish peroxidase conjugated protein A as an enzyme-labelled complex and a dilution by 100 folds of IgG from antiserum of rabbit to PRV-Ys strain as the working concentration. and was found to b workable for assessing an amount of PRV particles at ng level (ave.10ng/spot, at 37℃). A PRV-RNA cDNA molecular probe (the insert ca. 300bp) was successfully constructed wit...