کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8495949 | 1552914 | 2011 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Improved immunodetection of Penaeus monodon densovirus with monoclonal antibodies raised against recombinant capsid protein
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
علوم کشاورزی و بیولوژیک
علوم آبزیان
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Penaeus monodon densovirus (PmDNV), also called hepatopancreatic parvovirus (HPV), is a pathogen that causes stunt disease in the tiger shrimp, Penaeus monodon. In this study, the gene sequence encoding the capsid protein of PmDNV was cloned into two parts: PmDNV-N (837 bp) and PmDNV-C (806 bp). Both PCR products were cloned into the pGEX-6P-1 expression vector and transformed into the Escherichia coli BL21 strain. After induction, glutathione-S-transferase (GST)-tagged PmDNV-N and GST-tagged PmDNV-C proteins were obtained with molecular masses of 57.2 and 57.6 kDa, respectively. The recombinant proteins were purified by SDS-PAGE and used to immunize mice for monoclonal antibody (MAb) production. Three hybridoma clones producing MAbs specific to PmDNV-N and two hybridoma clones producing MAbs specific to PmDNV-C were selected by dot blot, Western blot and immunohistochemistry. All five MAbs detect PmDNV infection in shrimp by dot blot and Western blot, but only PmDNV-specific MAbs can be used to detect PmDNV infection in the hepatopancreas by immunohistochemistry. The detection limit of PmDNV-N-specific MAbs obtained in this study is similar to the most sensitive MAb characterized in a previous study, approximately 50 fmol μlâ 1 of antigen as determined by dot blot; the sensitivity of PmDNV-C-specific MAbs is twofold less than the MAbs specific to PmDNV-N. Using three MAbs specific for PmDNV-N and PmDNV-C together is twofold more sensitive than using either MAb alone. Although the detection limit of combining MAbs was 25,000 times lower than one-step PCR-based methods, using these MAbs is still useful for confirmation of PmDNV infection which is more convenient than normal histopathology confirmation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Aquaculture - Volume 311, Issues 1â4, 3 February 2011, Pages 19-24
Journal: Aquaculture - Volume 311, Issues 1â4, 3 February 2011, Pages 19-24
نویسندگان
Chutima Srisuk, Parin Chaivisuthangkura, Wasana Sukhumsirichart, Pattarin Sridulyakul, Siwaporn Longyant, Sombat Rukpratanporn, Paisarn Sithigorngul,