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Bui Thi Bich Hang. Development of a monoclonal antibody assay for Infectious Hypodermal and Hematopoietic Necrois Virus (IHHINV) of shrimp . Master's Degree(Biotechnology). Mahidol University. : Mahidol University, 2007.
Development of a monoclonal antibody assay for Infectious Hypodermal and Hematopoietic Necrois Virus (IHHINV) of shrimp
Name: Bui Thi Bich Hang
MeSH:
Blotting, Western
MeSH:
Densovirinae
MeSH:
Immunoblotting
MeSH:
Immunohistochemistry
Abstract:
The IHHN virus has been recognized as a disease of penaeid shrimp since 1981 in the
imported blue shrimp Penaeus stylirostris in Hawaii. It then was found to have spread
widely to Tahiti, Florida, Texas, Cayman Islands, Israel, Panama, Costa Rica, Belize,
Ecuador, Philippines, Singapore, Guam, Brazil, Honduras, France, Jamaica, and
recently in Thailand. Consequences of IHHNV infection in shrimp cultures were
assessed to cause serious problems for shrimp production and its values. Several
methods including histological examination, in situ hybridization and PCR are used to
detect IHHNV infection, but these methods are time consuming and expensive. This
study, therefore, developed monoclonal antibodies as one of the most rapid, simple,
accurate, and low cost methods to test the virus infection. Gene encoding GP3 capsid
protein of IHHNV was amplified and constructed into pET15b plasmid and
transformed into Escherichia coli BL21. Recombinant GP3 capsid protein of 37 kDa
was then expressed and extracted for immunization of mice (5 female BALB/C mice).
The polyclonal antibody was collected after 4 injections of antigen into the mice and
tested by western blot and immunohistochemistry. The mouse IV gave a strong signal
with IHHNV inclusion and was chosen for producing the monoclonal antibody
(MAb). The MAb was tested by dot blot ELISA, western blot as well as
immunohistochemistry. By dot blot ELISA, the monoclonal antibody detected GP3
capsid protein with different sensitivities ranging from 0.1 – 0.5 x 10-3 μg/μl. By
western blot, MAb-1 could detect a 37 kDa protein in the lysate of E. coli containing
GP3-pET15b at a dilution of 1: 100 in skimmed milk. The result of
immunohistochemistry showed that MAb strongly bound to IHHNV inclusions in the
gill, nerve and epithelial cells of P. vannamei infected with IHHNV. MAb were also
tested against other shrimp viral pathogens (HPV, WSSV, YHV and MBV) using
immunohistochemistry, and the results showed no cross-reactivity of the antibodies to
tested viruses.
Mahidol University
Address:
NAKHON PATHOM
Email:
liwww@mahidol.ac.th
Name: Manop Suphantharika
Role:
Thesis Advisors
Created:
2007
Modified:
2553-06-25
Issued:
2010-06-08
วิทยานิพนธ์/Thesis
application/pdf
CallNumber:
TH B933d 2007
eng
DegreeName: Master of Science
Level: Master's Degree
Descipline: Biotechnology
Grantor: Mahidol University
©copyrights Mahidol University
RightsAccess:
ใช้เวลา
0.277069 วินาที
Bui Thi Bich Hang
| Title | Contributor | Type |
|---|---|---|
| Development of a monoclonal antibody assay for Infectious Hypodermal and Hematopoietic Necrois Virus (IHHINV) of shrimp
มหาวิทยาลัยมหิดล Bui Thi Bich Hang | Manop Suphantharika | วิทยานิพนธ์/Thesis |
Manop Suphantharika