title: Detection of shrimp Taura syndrome virus by loop-mediated isothermal amplification using a designed portable multi-channel turbidimeter.
abstract: In this study, a portable turbidimetric end-point detection method was devised and tested for the detection of Taura syndrome virus (TSV) using spectroscopic measurement of a loop-mediated isothermal amplification (LAMP) by-product: magnesium pyrophosphate (Mg(2)P(2)O(7)). The device incorporated a heating block that maintained an optimal temperature of 63°C for the duration of the RT-LAMP reaction. Turbidity of the RT-LAMP by-product was measured when light from a light-emitting diode (LED) passed through the tube to reach a light dependent resistance (LDR) detector. Results revealed that turbidity measurement of the RT-LAMP reactions using this device provided the same detection sensitivity as the agarose gel electrophoresis detection of RT-LAMP and nested RT-PCR (IQ2000™) products. Cross reactions with other shrimp viruses were not found, indicating that the RT-LAMP-turbidity measurement was highly specific to TSV. The combination of 10 min for rapid RNA preparation with 30 min for RT-LAMP amplification followed by turbidity measurement resulted in a total assay time of less than 1h compared to 4-8h for the nested RT-PCR method. RT-LAMP plus turbidity measurement constitutes a platform for the development of more rapid and user-friendly detection of TSV in the field.
authors.affiliation: National Electronics and Computer Technology Center, 112 Pahol Yothin Rd., Klong Luang, Pathumthani 12120, Thailand.
authors.forename: Assawapong
authors.initials: A
authors.lastname: Sappat
authors.forename: Wansadaj
authors.initials: W
authors.lastname: Jaroenram
authors.forename: Teeranart
authors.initials: T
authors.lastname: Puthawibool
authors.forename: Tanom
authors.initials: T
authors.lastname: Lomas
authors.forename: Adisorn
authors.initials: A
authors.lastname: Tuantranont
authors.forename: Wansika
authors.initials: W
authors.lastname: Kiatpathomchai
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D056629:Dicistroviridae / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D009391:Nephelometry and Turbidimetry / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D033561:Penaeidae / Q000821:virology*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers