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• Chen, Z
• Liu, J
• Zeng, M
• Wang, Z
• Yu, D
• Yin, C
• Jin, L
• Yang, S
• Song, B
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• Chen, Z
• Liu, J
• Zeng, M
• Wang, Z
• Yu, D
• Yin, C
• Jin, L
• Yang, S
• Song, B
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• Liu, J
• Zeng, M
• Wang, Z
• Yu, D
• Yin, C
• Jin, L
• Yang, S
• Song, B

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Molecules 2012, 17(6), 6886-6900; doi:10.3390/molecules17066886

Article

Dot Immunobinding Assay Method with Chlorophyll Removal for the Detection of Southern Rice Black-Streaked Dwarf Virus

Zhuo Chen ¹ , Jiaju Liu ¹ , Mengjiao Zeng ² , Zhenchao Wang ¹ , Dandan Yu ¹ , Chengjun Yin ¹ , Linhong Jin ¹ , Song Yang ^1,*  and Baoan Song ^1,*

¹ State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang 550025, China ² Key Laboratory of Green Pesticide and Agriculture Bioengineering, Ministry of Education, Guizhou University, Guiyang 550025, China

* Authors to whom correspondence should be addressed.

Received: 23 April 2012; in revised form: 30 May 2012 / Accepted: 31 May 2012 / Published: 5 June 2012

Download PDF Full-Text [443 KB, uploaded 5 June 2012 10:28 CEST]

Abstract: Southern rice black-streaked dwarf virus (SRBSDV), a new virus from Fiji, has seriously damaged rice crops in southern China and northern Vietnam in recent years. This virus is difficult to diagnose in the early stages of infection, and is very destructive at the late stage. In the present study, a dot immunobinding assay (DIBA) that has a high sensitivity for diagnosing SRBSDV was developed. Two kinds of treatment for the DIBA were evaluated to determine the most effective one for removing chlorophyll interferences via rice extraction. The first included several reagents to remove chlorophyll, namely, the alkaline reagents like magnesium oxide and alumina oxide, the adsorbent reagents like activated carbon and bentonite, as well as the extraction agent acetone. The second and third treatments, which were used to remove chlorophyll in blot membrane-nitrocellulose and polyvinylidene fluoride (PVDF), included several organic solvents containing methanol, ethanol, acetone, ethyl acetate, and diethyl ether. The results showed that activated carbon and methanol yielded the best contrasting purple color for the infected samples by decreasing the chlorophyll content.

Keywords: Southern rice black-streaked dwarf virus (SRBSDV); dot immunobinding assay (DIBA); chlorophyll removal

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