The application of immunoblotting to the phenotyping of group-specific component |
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| Authors: | A S Thomas A J Ansford J G Aaskov |
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| Affiliation: | 1. State Health Laboratory, 63 George Street, Brisbane 4000, Australia;2. Department of Medical Laboratory Science, Queensland Institute of Technology, George Street, Brisbane 4000, Australia;1. Department of Chemical Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, UKM Bangi, Selangor, Malaysia;2. Fuel Cell Institute, Universiti Kebangsaan Malaysia, 43600, UKM Bangi, Selangor, Malaysia;3. Centre for Nanomaterial Research, Institute of Science, Universiti Teknologi MARA, 40450, Shah Alam, Selangor, Malaysia;4. School of Chemistry and Environment, Faculty of Applied Sciences, Universiti Teknologi MARA, 40450, Shah Alam, Selangor, Malaysia;1. West China–Washington Mitochondria and Metabolism Center and Department of Anesthesiology, West China Hospital, Sichuan University, Chengdu 610041, Sichuan, PRC;2. Mitochondria and Metabolism Center, Department of Anesthesiology and Pain Medicine, University of Washington, Seattle, WA 98109, USA;3. Division of Molecular Medicine, Department of Anesthesiology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA;1. Department of Vascular Surgery, the Second Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230601, China;2. Department of Pathology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430014, China;3. Department of Rheumatology, the Central Hospital of Wuhan, Wuhan, Hubei, 430014, China;4. Nursing Department, Xiangyang No.1 People’s Hospital, Hubei University of Medicine, Xiangyang, Hubei, 441000, China;5. Faculty of Chemical Engineering Technology, Universiti Malaysia Perlis (UniMAP), Perlis, Malaysia;6. Institute of Nano Electronic Engineering, 01000 Kangar, Universiti Malaysia Perlis, Perlis, Malaysia;7. Department of Vascular Surgery, Changzhou No.2 People''s Hospital Affiliated to Nanjing Medical University, Changzhou, Jiangsu, 213000, China |
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| Abstract: | A sensitive immunoblotting method for the routine detection of group-specific component (GC) from fresh serum, and from control and casework bloodstains has been developed. GC phenotypes were separated in a thin layer polyacrylamide gel by isoelectric focusing, transferred to nitrocellulose by a rapid capillary blotting procedure, and detected using a double antibody enzyme immunoassay. This method is capable of phenotyping 8 ng of GC extracted from bloodstains, a four-fold increase in sensitivity when compared to immunofixation and silverstaining. A total of 2424 casework bloodstains have been analysed and GC phenotypes identified in 78% of samples. The method is suitable for use in routine laboratories and is more sensitive than other methods for GC phenotyping of casework bloodstains. |
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