20887752
not annotated - annotated - LINNAEUS only
Rapid and sensitive detection of infectious hypodermal and hematopoietic necrosis virus by loop-mediated isothermal amplification combined with a lateral flow dipstick.
Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is an important shrimp pathogen that causes mortality in Penaeus stylirostris and stunting (called runt deformity syndrome or RDS) in Penaeus vannamei. Loop-mediated isothermal amplification (LAMP) allows rapid amplification of nucleic acids under isothermal conditions. It can be combined with a chromatographic lateral flow dipstick (LFD) for highly specific, rapid and simple visual detection of IHHNV-specific amplicons. Using this protocol, a 30-min amplification followed by 5 min hybridization with an FITC-labeled DNA probe and 5 min LFD resulted in visualization of DNA amplicons trapped at the LFD test line. Thus, 10 min for rapid DNA extraction followed by LAMP combined with LFD detection resulted in a total assay time of approximately 50 min. Detection sensitivity was comparable to other methods used commonly for nested PCR detection of IHHNV but had the additional advantages of reduced assay time, confirmation of amplicon identity by hybridization and elimination of electrophoresis with carcinogenic ethidium bromide.
Ann file
T1 Species 33 87 infectious hypodermal and hematopoietic necrosis virus
T2 Species 171 225 Infectious hypodermal and hematopoietic necrosis virus
T3 Species 227 232 IHHNV
T4 Species 291 311 Penaeus stylirostris
N4 Reference T4 Taxonomy:29019
T5 Species 368 384 Penaeus vannamei
N5 Reference T5 Taxonomy:6689
T6 Species 633 638 IHHNV
T7 Species 1088 1093 IHHNV
N8 Reference T1 Taxonomy:1513224 Decapod penstyldensovirus 1
N1 Reference T2 Taxonomy:1513224 Decapod penstyldensovirus 1
N2 Reference T3 Taxonomy:1513224 Decapod penstyldensovirus 1
N3 Reference T6 Taxonomy:1513224 Decapod penstyldensovirus 1
N6 Reference T7 Taxonomy:1513224 Decapod penstyldensovirus 1