title: Rapid and sensitive detection of Laem-Singh virus by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick.
abstract: Laem-Singh virus (LSNV) was discovered recently in Thailand in farmed Giant Tiger shrimp (Penaeus monodon) displaying signs of slow growth syndrome. Loop-mediated isothermal amplification (LAMP) allows DNA to be amplified rapidly at a constant temperature. Here a reverse transcription (RT)-LAMP method was combined with a chromatographic lateral-flow dipstick (LFD) to detect LSNV RNA rapidly and specifically. The reaction was optimized at 65°C for 30 min and amplified DNA hybridized to an FITC-labeled oligonucleotide probe for 5 min was detected at LFD test line 5 min after application. Including 10 min for rapid RNA extraction, test results could be generated within 1h and did not require electrophoresis. Compared to an existing RT-PCR method, the RT-LAMP-LFD was also ∼1000-fold more sensitive in detecting LSNV RNA.
authors.affiliation: Centex Shrimp, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand.
authors.forename: Narong
authors.initials: N
authors.lastname: Arunrut
authors.forename: Yortyot
authors.initials: Y
authors.lastname: Seetang-Nun
authors.forename: Jurairat
authors.initials: J
authors.lastname: Phromjai
authors.forename: Wattana
authors.initials: W
authors.lastname: Panphut
authors.forename: Wansika
authors.initials: W
authors.lastname: Kiatpathomchai
mesh_terms: D000818:Animals; D005814:Genes, Viral / Q000235:genetics; D009693:Nucleic Acid Hybridization; D033561:Penaeidae / Q000821:virology*; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013696:Temperature