สืบค้นงานวิจัย
In situ DIG-labeling, loop-mediated DNA Amplification (ISDL) for highly sensitive detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV)
Jitrakorn S. - ไม่ระบุหน่วยงาน
ชื่อเรื่อง (EN): In situ DIG-labeling, loop-mediated DNA Amplification (ISDL) for highly sensitive detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV)
ผู้แต่ง / หัวหน้าโครงการ (EN): Jitrakorn S.
บทคัดย่อ (EN): Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is also known as Penaeus stylirostris densovirus (PstDNV). Since it is listed as a reportable crustacean disease by the World Organization for Animal Health (OIE), many countries require testing for IHHNV in shrimp stocks imported for aquaculture, necessitating rapid and highly sensitive detection methods to guarantee and maintain absence of IHHNV in traded stocks. Here we describe a new, highly sensitive method for detection of IHHNV in shrimp tissues. It involves in situ loop-mediated DNA amplification (IS-LAMP) employing digoxigenin-11-dUTP (DIG-dUTP) as part of the deoxynucleotide triphosphate (dNTP) mix. This resulted in DIG-labeling of the DNA amplicons (i.e., in situ, DIG-labeling LAMP or ISDL) allowing direct in situ, immuno-histochemical detection of IHHNV using an anti-DIG antibody conjugated with alkaline phosphatase, and eliminating the lengthy hybridization step of conventional ISH. The target region for the method is at the 3'end of the IHHNV genome that has been reported least likely to give rise to endogenous viral elements (EVE) in the shrimp genome. When compared to the results for IHHN detection by conventional in situ hybridization (ISH) using a DIG-labeled oligonucleotide probe, signals from the ISDL method were stronger and also gave positive results with some tissues unreactive by the conventional ISH method. Additional comparison of the two methods revealed that the proteinase-K digestion step needed for normal ISH was unnecessary for ISDL. This further shortened the assay time and improved the tissue integrity necessary to clearly indicate the location of positive hybridization signals. The ISDL method gave no cross- reactions with tissues of shrimp infected with other common pathogens, and the total time needed to carry out ISDL was one-third of that needed for conventional ISH. Statement of relevance: This work describes an in situ, DIG-labeling LAMP (ISDL) method, as a new, highly sensitive method for detection of IHHNV in shrimp tissues. The developed method can be used as a confirmatory test carried out with tissue sections to confirm the results from molecular assays, especially when immune-histochemical methods are not available to do so. ISDL is especially important for molecular detection from a new or unexpected location and for molecular detection in newly suspected carriers to determine whether they are actually infected or just mechanical carriers. © 2016 Elsevier B.V.
บทคัดย่อ: ไม่พบข้อมูลจากหน่วยงานต้นทาง
ภาษา (EN): en
เอกสารแนบ (EN): https://www.scopus.com/inward/record.uri?eid=2-s2.0-84960464009&doi=10.1016%2fj.aquaculture.2016.01.023&partnerID=40&md5=58e3cfeb7504be276554bb2cd5954bae
เผยแพร่โดย (EN): มหาวิทยาลัยมหิดล
คำสำคัญ (EN): Litopenaeus stylirostris
เจ้าของลิขสิทธิ์ (EN): มหาวิทยาลัยมหิดล
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การอ้างอิง

Jitrakorn S.. "In situ DIG-labeling, loop-mediated DNA Amplification (ISDL) for highly sensitive detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV)". ไม่ระบุหน่วยงาน. ม.ป.ป.

Jitrakorn S.. "In situ DIG-labeling, loop-mediated DNA Amplification (ISDL) for highly sensitive detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV)". ไม่ระบุหน่วยงาน. ม.ป.ป.. Print.



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In situ DIG-labeling, loop-mediated DNA Amplification (ISDL) for highly sensitive detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV)
Jitrakorn S.
มหาวิทยาลัยมหิดล
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Sensitive visual detection of AHPND bacteria using loop-mediated isothermal amplification combined with DNAFunctionalized gold nanoparticles as probes การวิเคราะห์จีโนมของกุ้งกุลาดำโดยใช้เทคนิค southern blot hybridization เพื่อประโยชน์ในการวินิจฉัยโรค infectious hypodermal and hematopoietic necrosis virus (IHHNV) Nanocontainer designed from an infectious hypodermal and hematopoietic necrosis virus (IHHNV) has excellent physical stability and ability to deliver shrimp tissues Sensitivity improvement of immunochromatographic strip test for infectious myonecrosis virus detection การพัฒนาวิธีการตรวจโรคเมลิออยโดสิสด้วย loop-mediated isothermal amplification (LAMP) Development of loop-mediated isothermal amplification (LAMP) for diagnosing melioidosis Development of reverse transcription-loop-mediated isothermal amplification (RT-LAMP) for the detection of the causative agent of covid-19 Infectious hypodermal and hematopoietic necrosis virus-like particles encapsulating VP28 double-stranded RNA protect shrimp from white spot syndrome virus การพัฒนาวิธีการตรวจโรคเมลิออยโดสิสและเลปโตสไปโรสิสด้วย loop-mediated isothermal amplification (LAMP) Development of loop-mediated isothermal amplification (LAMP) for diagnosing melioidosis and leptosp The performance of in-house loop-mediated isothermal amplification for rapid detection of Mycobacterium tuberculosis in sputum sample comparing to Xpert MTB/RIF, microscopy and culture Pragmatic accuracy of an in-house loop-mediated isothermal amplification (LAMP) for diagnosis of pulmonary tuberculosis in a Thai community hospital
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