Zhe Wang1, Yan Tang1,2, Hongwei Dang3, Xuxuan Zhang1, Lijuan Pang1, Pengyan Wang4, Chuangfu Chen4, Yan Ren1. 1. Department of Pathology and Key Laboratory of Xinjiang Endemic and Ethnic Diseases (Ministry of Education)/Department of Pathology, The First Affiliated Hospital, Shihezi University School of Medicine Shihezi, China. 2. Animal Science and Technology Branch, Xinjiang Agricultural Vocational Technical College Changji 831100, Xinjiang, China. 3. Department of Obstetrics and Gynecology, The First Affiliated Hospital of Shihezi University Shihezi 832000, Xinjiang, China. 4. College of Animal Science and Technology, Shihezi University Shihezi 832000, Xinjiang, China.
Abstract
BACKGROUND: Xinjiang, China shows the world's highest incidence and mortality rates of cervical cancer. Due to limited conditions available for medical examination, hybrid capture 2 (HC2) and other detection methods are used rarely, and early screening for human papillomavirus (HPV) cannot be carried out. Therefore, we established a double-antibody sandwich (DAS)-enzyme-linked immunosorbent assay (ELISA) based on a polymorphism of the Xinjiang HPV16 L1 strain (KU721788). METHODS: According to the conserved sequence and specific epitope of Xinjiang strain HPV16 L1, we prepared two anti-HPV16 L1 monoclonal antibodies and combined them to construct a DAS-ELISA. Detection conditions for the DAS-ELISA were optimized, and HC2 was used as the control to verify the specificity, repeatability and coincidence detection of the DAS-ELISA. RESULTS: The optimized conditions for the DAS-ELISA were: dilution of the capture antibody was 1:100; the enzyme-labelled antibody was 1:10; the sample reaction time was 45 min; the enzyme-labelled antibody was applied for 40 min, and the substrate color development time was 15 min. The quality of the DAS-ELISA for the detection of HPV 16 was very high, and there was no significant difference when compared with HC2. CONCLUSION: The DAS-ELISA developed on the basis of the Xinjiang strain (KU721788) polymorphism possesses the advantages of a detection rate similar to that for the HC2 assay currently used clinically, but it is more convenient operationally and at lower cost. DAS-ELISA is thus easier to implement for cervical cancer screening in economically depressed areas. IJCEP
BACKGROUND: Xinjiang, China shows the world's highest incidence and mortality rates of cervical cancer. Due to limited conditions available for medical examination, hybrid capture 2 (HC2) and other detection methods are used rarely, and early screening for human papillomavirus (HPV) cannot be carried out. Therefore, we established a double-antibody sandwich (DAS)-enzyme-linked immunosorbent assay (ELISA) based on a polymorphism of the Xinjiang HPV16 L1 strain (KU721788). METHODS: According to the conserved sequence and specific epitope of Xinjiang strain HPV16 L1, we prepared two anti-HPV16 L1 monoclonal antibodies and combined them to construct a DAS-ELISA. Detection conditions for the DAS-ELISA were optimized, and HC2 was used as the control to verify the specificity, repeatability and coincidence detection of the DAS-ELISA. RESULTS: The optimized conditions for the DAS-ELISA were: dilution of the capture antibody was 1:100; the enzyme-labelled antibody was 1:10; the sample reaction time was 45 min; the enzyme-labelled antibody was applied for 40 min, and the substrate color development time was 15 min. The quality of the DAS-ELISA for the detection of HPV 16 was very high, and there was no significant difference when compared with HC2. CONCLUSION: The DAS-ELISA developed on the basis of the Xinjiang strain (KU721788) polymorphism possesses the advantages of a detection rate similar to that for the HC2 assay currently used clinically, but it is more convenient operationally and at lower cost. DAS-ELISA is thus easier to implement for cervical cancer screening in economically depressed areas. IJCEP
Authors: F Nahar; M A Hossain; S K Paul; M U Ahmed; S Khatun; G R Bhuiyan; S A Nasreen; N Haque; S Ahmed; N Kobayashi; S N Akter; H Begum Journal: Mymensingh Med J Date: 2019-01