title: Highly sensitive serological methods for detecting tomato yellow leaf curl virus in tomato plants and whiteflies.
abstract: BACKGROUND
  Tomato yellow leaf curl virus (TYLCV) is a member of the genus Begomovirus in the family Geminiviridae, which causes severe losses in tomato production in tropic and subtropic regions.
  
  
  METHODS
  The purified TYLCV virions were used as the immunogen to produce monoclonal antibodies (MAbs) using the hybridoma technology. MAb-based dot enzyme-linked immunosorbent assay (dot-ELISA) and direct tissue blot immunoassay (DTBIA) were developed for sensitive, simple, and rapid detection of TYLCV in field tomato and whitefly (Bemisia tabaci) samples collected from TYLCV prevalent provinces in China.
  
  
  RESULTS
  Using the hybridoma technology, six murine MAbs (1C4, 8D10, 6E3, 2F2, 3F4 and 4G3) against TYLCV were prepared. Using the MAb 1C4, dot-ELISA and DTBIA were then established for detecting TYLCV in field tomato and whitefly samples collected from TYLCV prevalent provinces in China. The dot-ELISA could detect TYLCV in infected tissue crude extract diluted at 1:5,120 (w/v, g mL-1), and in viruliferous whitefly homogenate diluted at 1:128 (individual whitefly/μL), respectively. Field tomato samples (n=487) and whitefly samples (n=110) from TYLCV prevalent districts in China were screened for the presence of TYLCV using the two developed methods, and the results were further confirmed by PCR and nucleotide sequencing. The survey revealed that TYLCV is widespread on tomato plants in Zhejiang, Shandong and Henan provinces in China.
  
  
  CONCLUSIONS
  The developed dot-ELISA is very suitable for the routine detection of TYLCV in field tomato and whitefly samples, and the DTBIA is more suitable for the routine detection of TYLCV in large-scale tomato plant samples collected from TYLCV prevalent areas.
authors: State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou, Zhejiang 310058, China.
authors: Yan
authors: Y
authors: Xie
authors: Xiaoyang
authors: X
authors: Jiao
authors: Xueping
authors: X
authors: Zhou
authors: Huan
authors: H
authors: Liu
authors: Yuequn
authors: Y
authors: Ni
authors: Jianxiang
authors: J
authors: Wu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000302:isolation & purification; D053839:Begomovirus / Q000302:isolation & purification*; D006430:Hemiptera / Q000821:virology*; D007118:Immunoassay / Q000379:methods; D018551:Solanum lycopersicum / Q000821:virology*; D051379:Mice; D010935:Plant Diseases / Q000821:virology*; D013997:Time Factors; D014773:Virology / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral
references: Virol J. 2011 May 15;8:228
references: 21569615
references: Plant Dis. 1999 May;83(5):487
references: 30845549
references: Virus Genes. 2009 Oct;39(2):249-55
references: 19590945
references: Virology. 1991 Nov;185(1):151-61
references: 1926771
references: Plant Dis. 1999 Nov;83(11):984-988
references: 30841296
references: Nucleic Acids Res. 1991 Dec 25;19(24):6763-9
references: 1840676
references: PLoS Pathog. 2010 Oct 28;6(10):e1001164
references: 21060815
references: Plant Dis. 2006 Oct;90(10):1359
references: 30780951
references: Phytopathology. 2005 Sep;95(9):1089-97
references: 18943307
references: Plant Dis. 2001 Feb;85(2):230
references: 30831958