ABSTRACT
The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21-CodonPlus (DE3)-RIL using the pGEX-4T-1 expression vector and the soluble recombinant product was purified with Glutathione Sepharose TM<'4B> by centrifugation. The soluble recombinant protein showed good immune reactions and was confirmed by Western blot using anti-CSFV-specific antibodies. Then an indirect ELISA with the purified E2 protein as the coating antigen was established to detect antibody against CSFV. The result revealed that the optimal concentration of coated antigen was 0.6 μg/well and the optimal dilution of serum was 1:80. The positive cut-off value of this ELISA assay was OD<,tested serum>/OD<,negative serum>≥2.1- The E2-ELISA method was evaluated by comparison with the indirect hemagglutination test (IHAT). When a total of 100 field serum samples were tested the sensitivity and specificity were 90.3% and 94.7% respectively. Specificity analysis showed that there were no cross-reactions between BVD serum and the purified E2 protein in the E2-ELISA.
ABSTRACT
Background: \r\n', u'Chicken embryo cell was the most effective fibroblast cell line for producing the attenuated measles vaccines. Many countries as Japan, in Kitasato, an attenuated virus derived from the Edmonston virulent strain adapted in chick embryo cells, it was call AIK- C strain, and used for measles vaccine production. In Vietnam, the POLYVAC have applied the Kitasato institute\u2019s technology into measles vaccines production. The Valo SPF egg (Lohmann-Germany) was used in the chick embryo culture \r\n', u'Objectives: \r\n', u'To study the inoculation, and clone of the human AIK-C strain in the chick embryo in order to achieve a high yield of virus for measles vaccine production \r\n', u'Subjects and method:\r\n', u'Suitablecells (AIK- C Strain, an attenuated virus derived from the Edmonston virulent strain, chicken embryo cell cultured from SPF egg Valo Germany) are selected. Then methods as inoculating and culturinglive viruses on the chicken embryo cell are applied.\r\n', u'Results:\r\n', u'After 9 days for infection, viruses which were observed via microscope, propagated and in infectious clone by the measles virus.\r\n', u'Conclusion: \r\n', u'Many tests for propagation of AIK measles virus in chicken embryo cell cuture derived from Valo SPF egg (Germany) showed that chicken embryo cell culture from Valo SPF egg (Germany) is appropriated fro AIK- C measles vaccine production in Vietnam.\r\n', u'
Subject(s)
Measles virusABSTRACT
BACKGROUND: Human herpesvirus 8 (HHV8) has been detected in all forms of Kaposi's sarcoma (KS). The role of HHV 8 in dermatologic diseases other than KS is controversial. Some studies based on polymerase chain reaction (PCR) findings suggest an association between HHV8 and epithelial tumors of the skin. OBJECTIVE: To assess the presence of HHV8 sequences in classic KS and some premalignant and malignant cutaneous epithelial tumors by nested PCR and find out any variations in HHV8 DNA by direct sequencing. METHODS: In total 71 sample tissues were obtained from Korean patients: 5 classic KS, 12 Bowen's disease (BD), 19 actinic keratosis, 14 squamous cell carcinomas and 21 basal cell carcinomas. The first and nested PCR were performed to amplify the fragments of HHV8 open read frame (ORF) 26 and ORF 75 which were directly sequenced by dideoxy termination methods. RESULTS: HHV8 DNA sequences were found in 3 of 5 classic KS (60%) and in 1 of 12 BD. Other epithelial tumors were negative for HHV8. On sequencing PCR products, substitutions at 1033 C->T, 1086 C->T, and 1139 A->C were detected in ORF 26 of all 4 positive PCR products. In 2 of 3 KS positive for ORF 26, substitution was detected at 237 G->A in ORF 75. CONCLUSION: All of our Korean samples were included in the Zong's C strain. This study suggests that C strain is predominant in classic KS in Korea. But it is rare in non-KS cutaneous epithelial tumors. Studies of the length difference in the HHV8 genome should follow.
Subject(s)
Animals , Humans , Base Sequence , Bowen's Disease , Carcinoma, Basal Cell , Carcinoma, Squamous Cell , DNA , Ecthyma, Contagious , Genome , Herpesvirus 8, Human , Keratosis, Actinic , Korea , Polymerase Chain Reaction , Sarcoma, Kaposi , SkinABSTRACT
BACKGROUND: Human herpesvirus 8 (HHV8) has been detected in all forms of Kaposi's sarcoma (KS). The role of HHV 8 in dermatologic diseases other than KS is controversial. Some studies based on polymerase chain reaction (PCR) findings suggest an association between HHV8 and epithelial tumors of the skin. OBJECTIVE: To assess the presence of HHV8 sequences in classic KS and some premalignant and malignant cutaneous epithelial tumors by nested PCR and find out any variations in HHV8 DNA by direct sequencing. METHODS: In total 71 sample tissues were obtained from Korean patients: 5 classic KS, 12 Bowen's disease (BD), 19 actinic keratosis, 14 squamous cell carcinomas and 21 basal cell carcinomas. The first and nested PCR were performed to amplify the fragments of HHV8 open read frame (ORF) 26 and ORF 75 which were directly sequenced by dideoxy termination methods. RESULTS: HHV8 DNA sequences were found in 3 of 5 classic KS (60%) and in 1 of 12 BD. Other epithelial tumors were negative for HHV8. On sequencing PCR products, substitutions at 1033 C->T, 1086 C->T, and 1139 A->C were detected in ORF 26 of all 4 positive PCR products. In 2 of 3 KS positive for ORF 26, substitution was detected at 237 G->A in ORF 75. CONCLUSION: All of our Korean samples were included in the Zong's C strain. This study suggests that C strain is predominant in classic KS in Korea. But it is rare in non-KS cutaneous epithelial tumors. Studies of the length difference in the HHV8 genome should follow.