title: Development of loop-mediated isothermal amplification and SYBR green real-time PCR methods for the detection of Citrus yellow mosaic badnavirus in citrus species.
abstract: Citrus yellow mosaic badnavirus (CMBV) is an important pathogen in southern India spread by infected citrus propagules. One of the measures to arrest the spread of CMBV is to develop methods to screen and certify citrus propagules as CMBV-free. The methods loop-mediated isothermal amplification (LAMP) and SYBR green real-time PCR (SGRTPCR) have been developed for the efficient detection of CMBV in citrus propagules. This paper compares the sensitivities of LAMP and SGRTPCR with polymerase chain reaction (PCR) for the detection of CMBV. Whereas PCR and LAMP were able to detect CMBV from a minimum of 10 ng of total DNA of infected leaf samples, SGRTPCR could detect the same from 1 ng of total DNA. Using SGRTPCR, the viral titres were estimated to be the highest in rough lemon and lowest in Nagpur Mandarin of the five naturally infected citrus species tested. The results will help in designing suitable strategies for the sensitive detection of CMBV from citrus propagules.
authors.affiliation: Department of Virology, Sri Venkateswara University, Tirupati 517502, Andhra Pradesh, India.
authors.forename: A M
authors.initials: AM
authors.lastname: Anthony Johnson
authors.affiliation: Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi 110021, India. Electronic address: indranil58@yahoo.co.in.
authors.forename: I
authors.initials: I
authors.lastname: Dasgupta
authors.affiliation: Department of Virology, Sri Venkateswara University, Tirupati 517502, Andhra Pradesh, India.
authors.forename: D V R
authors.initials: DV
authors.lastname: Sai Gopal
mesh_terms: D019195:Badnavirus / Q000235:genetics / Q000302:isolation & purification*; D052160:Benzothiazoles; D002957:Citrus / Q000821:virology*; D003959:Diamines; D007194:India; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D010935:Plant Diseases / Q000821:virology; D011804:Quinolines; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D013696:Temperature
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I
keywords: Badnavirus; Citrus; LAMP; PCR; Real-time PCR