Retracted: Reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with colorimetric gold nanoparticle (AuNP) probe assay for visual detection ofPenaeus vannameinodavirus (PvNV)

Abstract

Significance and Impact of the Study: This study represents the first report of Penaeus vannamei nodavirus (PvNV) detection using the colorimetric RT-LAMP technique. The application of RT-LAMP assay combined with colorimetric gold nanoparticle (AuNP) probe to detect the PvNV offers simple, rapid and sensitive technique and does not require sophisticated equipment with applicable for small or field laboratories. This method can further employ to screen broodstock before breeding, to screen postlarvae before ponds stocking, to monitor shrimp in rearing ponds and to assess the occurrence of this virus in shrimp farming. AbstractPenaeus vannamei nodavirus (PvNV) is an emerging viral infection that has caused muscle necrosis or a white tail disease in cultivated whiteleg shrimp Penaeus (Litopenaeus) vannamei, leading to loss in the productions. Rapid detection of PvNV is essential for further control disease. A combination between reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay and colorimetric gold nanoparticle (AuNP) probe was developed for rapid, sensitive and inexpensive detection of PvNV in this study. RT-LAMP reaction successfully detected PvNV at 63°C for 45 and 5 min for hybridization of LAMP amplified product, followed by salt-induced AuNPlabelled ssDNA probe aggregation to visual colour development. This method showed identical results to LAMP followed by gel electrophoresis and spectrophotometric detection, and it was 10 times sensitive more than conventional nested RT-PCR. The new method revealed negative results with other shrimp pathogens. This study provides the direct visualization of LAMP reaction by AuNP probe hybridization, that significantly reduces the time and cost required for the molecular diagnostic of infectious diseases and offers to use in aquaculture health management.

Rapid Colorimetric Assay for Detection of Listeria monocytogenes in Food Samples Using LAMP Formation of DNA Concatemers and Gold Nanoparticle-DNA Probe Complex

“…A ratio of the optimum hybridization condition was modified from Suebsing et al ( 2013 ). The hybridization for the detection of LAMP products was conducted in a total volume of 6 μL at 50°C for 10 min.…”

Advances in the study of nodavirus

“…Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) is another potential point-of-care diagnostic assay, as a laboratory setup such as thermocycler and electrophoresis equipment can be omitted. Suebsing, Prombun & Kiatpathomchai (2013) developed an RT-LAMP with colorimetric gold nanoparticle probe assay for the detection of PvNV in P. vannamei and P. monodon . This assay is 10x more sensitive than the nested RT-PCR established by Tang et al (2007) .…”

SEBARAN INFEKSI TAURA SYNDROME, INFECTIOUS MYONECROSIS, DAN Penaeus vannamei NERVOUS VIRUS (TSV, IMNV, DAN PvNV) PADA BUDIDAYA UDANG Litopenaeus vannamei DI JAWA BARAT, JAWA TIMUR, DAN BALI

“…Virus-virus tersebut dapat dideteksi dengan beberapa metode, seperti metode molekuler (reverse transcription-polymerase chain reaction(RT-PCR), nested RT-PCR, Real Time RT-PCR, in-situ hybridisation (ISH), dan Reverse transcription loop-mediated isothermal amplification (RT-LAMP) yang dikombinasikan dengan colorimetric gold nanoparticle (AuNP) (Tang et al, 2005;Senapin et al, 2007;Andrade et al, 2007;Suebsing et al, 2013 Setelah proses membalikan dari RNA ke c-DNA menggunakan reverse enzyme pada suhu 50°C-60°C serta 94°C, selanjutnya c-DNA diamplifikasi dengan suhu annealing yang berbeda untuk masing-masing target virus di mana TSV menggunakan suhu 60°C, IMNV 50°C, dan untuk PvNV 55°C. Selanjutnya sampel dielektrophoresis pada 1,5% agarose dalam larutan penyangga atau buffer 1x TAE (Triss Acetate EDTA).…”