Screening and identification of B cell epitope within the major capsid protein L1 of HPV 52, using monoclonal antibodies
In this study, HPV52 L1 protein was induced to be expressed. Monoclonal antibody (mAb) 6A7 against L1 protein were screened by cell fusion techniques. Western Blot and immunofluorescence assay (IFA) demonstrated the specificity of the mAb. The L1 protein was truncated for prokaryotic expression (N1~N7) and Dot-ELISA showed that 6A7 recognized N3 (aa 200~350). The immunodominant regions were truncated again for expression, with 6A7 recognizing N6 (aa 251~305). The N6 proteins were further truncated and then were constructed an four-segment eukaryotic expression vector. IFA showed that 6A7 could recognize amino acid 262~279....
Source: Journal of Virological Methods - November 28, 2023 Category: Virology Authors: Yumei Chen Shan Zhang Gaiping Zhang Jingming Zhou Hongliang Liu Chao Liang Enping Liu Xifang Zhu Aiping Wang Source Type: research
Screening and identification of B cell epitope within the major capsid protein L1 of HPV 52, using monoclonal antibodies
In this study, HPV52 L1 protein was induced to be expressed. Monoclonal antibody (mAb) 6A7 against L1 protein were screened by cell fusion techniques. Western Blot and immunofluorescence assay (IFA) demonstrated the specificity of the mAb. The L1 protein was truncated for prokaryotic expression (N1~N7) and Dot-ELISA showed that 6A7 recognized N3 (aa 200~350). The immunodominant regions were truncated again for expression, with 6A7 recognizing N6 (aa 251~305). The N6 proteins were further truncated and then were constructed an four-segment eukaryotic expression vector. IFA showed that 6A7 could recognize amino acid 262~279....
Source: Journal of Virological Methods - November 28, 2023 Category: Virology Authors: Yumei Chen Shan Zhang Gaiping Zhang Jingming Zhou Hongliang Liu Chao Liang Enping Liu Xifang Zhu Aiping Wang Source Type: research
Performance characteristcis of Allele-Specific PCR (ASPCR) in detecting drug resistance mutations among non-B HIV-1 Variants
J Virol Methods. 2023 Nov 22:114856. doi: 10.1016/j.jviromet.2023.114856. Online ahead of print.ABSTRACTAllele-Specific Polymerase Chain Reaction (ASPCR) is an affordable point-mutation assay whose validation could improve the detection of HIV-1 drug resistance mutations (DRMs) in resource-limited settings (RLS). We assessed the performance of ASPCR onforty-four non-B HIV-1 plasma samples from patients who were ARV treated in failure in N'Djamena-Chad. Viral RNA was reverse-transcribed and amplified using LightCycler® FastStart DNA MasterPLUS SYBR Green I. Detection of six major DRMs (K70R, K103N, Y181C, M184V, T215F, T21...
Source: Journal of Virological Methods - November 24, 2023 Category: Virology Authors: Chatt é Adawaye Joseph Fokam Erick Ntambwe Kamangu Derrick Tambe Ayuk Ngwese Fabrice Susin Ali Mahamat Moussa BertinTchombou Hig-Zounet Joseph Mad-Toingu é Abdelsalam Tidjani Dolores Vaira Michel Moutschen Source Type: research
Performance characteristcis of Allele-Specific PCR (ASPCR) in detecting drug resistance mutations among non-B HIV-1 Variants
J Virol Methods. 2023 Nov 22:114856. doi: 10.1016/j.jviromet.2023.114856. Online ahead of print.ABSTRACTAllele-Specific Polymerase Chain Reaction (ASPCR) is an affordable point-mutation assay whose validation could improve the detection of HIV-1 drug resistance mutations (DRMs) in resource-limited settings (RLS). We assessed the performance of ASPCR onforty-four non-B HIV-1 plasma samples from patients who were ARV treated in failure in N'Djamena-Chad. Viral RNA was reverse-transcribed and amplified using LightCycler® FastStart DNA MasterPLUS SYBR Green I. Detection of six major DRMs (K70R, K103N, Y181C, M184V, T215F, T21...
Source: Journal of Virological Methods - November 24, 2023 Category: Virology Authors: Chatt é Adawaye Joseph Fokam Erick Ntambwe Kamangu Derrick Tambe Ayuk Ngwese Fabrice Susin Ali Mahamat Moussa BertinTchombou Hig-Zounet Joseph Mad-Toingu é Abdelsalam Tidjani Dolores Vaira Michel Moutschen Source Type: research
Performance characteristcis of Allele-Specific PCR (ASPCR) in detecting drug resistance mutations among non-B HIV-1 Variants
J Virol Methods. 2023 Nov 22:114856. doi: 10.1016/j.jviromet.2023.114856. Online ahead of print.ABSTRACTAllele-Specific Polymerase Chain Reaction (ASPCR) is an affordable point-mutation assay whose validation could improve the detection of HIV-1 drug resistance mutations (DRMs) in resource-limited settings (RLS). We assessed the performance of ASPCR onforty-four non-B HIV-1 plasma samples from patients who were ARV treated in failure in N'Djamena-Chad. Viral RNA was reverse-transcribed and amplified using LightCycler® FastStart DNA MasterPLUS SYBR Green I. Detection of six major DRMs (K70R, K103N, Y181C, M184V, T215F, T21...
Source: Journal of Virological Methods - November 24, 2023 Category: Virology Authors: Chatt é Adawaye Joseph Fokam Erick Ntambwe Kamangu Derrick Tambe Ayuk Ngwese Fabrice Susin Ali Mahamat Moussa BertinTchombou Hig-Zounet Joseph Mad-Toingu é Abdelsalam Tidjani Dolores Vaira Michel Moutschen Source Type: research
A Sensitive Luciferase Reporter Assay for the Detection of Infectious African Swine Fever Virus
J Virol Methods. 2023 Nov 19:114854. doi: 10.1016/j.jviromet.2023.114854. Online ahead of print.ABSTRACTAfrican swine fever virus (ASFV) is a complex DNA virus causing severe hemorrhagic disease in domestic pigs and wild boar. The disease has spread worldwide, with important socio-economic consequences. Early virus detection and control measures are crucial as there are no effective vaccines nor antivirals on the market. While the diagnosis of ASFV is fast and based primarily on qPCR, the detection of infectious ASFV is a labor-intensive process requiring susceptible macrophages and subsequent antibody-based staining or he...
Source: Journal of Virological Methods - November 21, 2023 Category: Virology Authors: Kemal Mehinagic Matthias Liniger Maksym Samoilenko Nick Soltermann Markus Gerber Nicolas Ruggli Source Type: research
Comparison of Polymerase Chain Reaction (PCR) assay performance in detecting Decapod penstylhamaparvovirus 1 in penaeid shrimp
J Virol Methods. 2023 Nov 19:114840. doi: 10.1016/j.jviromet.2023.114840. Online ahead of print.ABSTRACTDecapod Penstylhamaparvovirus 1, commonly known as infectious hypodermal and hematopoietic necrosis virus (IHHNV), remains an economically important viral pathogen for penaeid shrimp aquaculture due to its effects on growth performance. The World Organization for Animal Health (WOAH, Paris, France) recommended methods for the detection of IHHNV include both conventional and real-time PCR. However, published reports and anecdotal evidence suggest the occurrence of non-specific amplifications when testing for IHHNV using t...
Source: Journal of Virological Methods - November 21, 2023 Category: Virology Authors: Arun K Dhar Roberto Cruz-Flores Hung N Mai Janet Warg Source Type: research
A Sensitive Luciferase Reporter Assay for the Detection of Infectious African Swine Fever Virus
J Virol Methods. 2023 Nov 19:114854. doi: 10.1016/j.jviromet.2023.114854. Online ahead of print.ABSTRACTAfrican swine fever virus (ASFV) is a complex DNA virus causing severe hemorrhagic disease in domestic pigs and wild boar. The disease has spread worldwide, with important socio-economic consequences. Early virus detection and control measures are crucial as there are no effective vaccines nor antivirals on the market. While the diagnosis of ASFV is fast and based primarily on qPCR, the detection of infectious ASFV is a labor-intensive process requiring susceptible macrophages and subsequent antibody-based staining or he...
Source: Journal of Virological Methods - November 21, 2023 Category: Virology Authors: Kemal Mehinagic Matthias Liniger Maksym Samoilenko Nick Soltermann Markus Gerber Nicolas Ruggli Source Type: research
Comparison of Polymerase Chain Reaction (PCR) assay performance in detecting Decapod penstylhamaparvovirus 1 in penaeid shrimp
J Virol Methods. 2023 Nov 19:114840. doi: 10.1016/j.jviromet.2023.114840. Online ahead of print.ABSTRACTDecapod Penstylhamaparvovirus 1, commonly known as infectious hypodermal and hematopoietic necrosis virus (IHHNV), remains an economically important viral pathogen for penaeid shrimp aquaculture due to its effects on growth performance. The World Organization for Animal Health (WOAH, Paris, France) recommended methods for the detection of IHHNV include both conventional and real-time PCR. However, published reports and anecdotal evidence suggest the occurrence of non-specific amplifications when testing for IHHNV using t...
Source: Journal of Virological Methods - November 21, 2023 Category: Virology Authors: Arun K Dhar Roberto Cruz-Flores Hung N Mai Janet Warg Source Type: research
Novel high-coverage primers for detection of canine morbillivirus by end-point and real-time RT-PCR assays
J Virol Methods. 2023 Nov 16:114853. doi: 10.1016/j.jviromet.2023.114853. Online ahead of print.ABSTRACTCanine distemper virus (CDV) is a major threat to domestic dogs and wildlife worldwide. Molecular assays are the most sensitive and specific tests to diagnose the disease, however, the high CDV genetic variability may compromise laboratory diagnosis. Herein, we designed a high-coverage primer set for end-point (RT-PCR) and real-time (RT-qPCR) for CDV detection. Initially, we collected 194 complete/near-complete CDV genomes (GenBank) and analyzed them for highly conserved regions for primer design. We then assessed the in...
Source: Journal of Virological Methods - November 18, 2023 Category: Virology Authors: Alice Silveira Becker Tha ísa Regina Rocha Lopes Nat ália Hettwer Pedroso Jos é Valter Joaquim Silva Júnior Rudi Weiblen Eduardo Furtado Flores Source Type: research
Purification of Hepatitis C Virus Core Protein in Non-Denaturing Condition
In this study, we successfully solubilized bacterially expressed core protein using a high-salt and detergent-containing buffer and bypassed the denaturing-refolding process. Size-exclusion chromatography revealed three distinct peaks in the HCV-infected cell lysate, with the bacterially expressed soluble core protein corresponding to its second peak. Using a combination of affinity, size exclusion, and multi-modal chromatography purification techniques, we achieved a purity of >95% for the core protein. Analytical ultracentrifugation revealed monomer formation in the solution. Far UV Circular dichroism spectroscopy ide...
Source: Journal of Virological Methods - November 18, 2023 Category: Virology Authors: Kyo Izumida Yumiko Hara Yukio Furukawa Kotaro Ishida Keisuke Tabata Eiji Morita Source Type: research
Novel high-coverage primers for detection of canine morbillivirus by end-point and real-time RT-PCR assays
J Virol Methods. 2023 Nov 16:114853. doi: 10.1016/j.jviromet.2023.114853. Online ahead of print.ABSTRACTCanine distemper virus (CDV) is a major threat to domestic dogs and wildlife worldwide. Molecular assays are the most sensitive and specific tests to diagnose the disease, however, the high CDV genetic variability may compromise laboratory diagnosis. Herein, we designed a high-coverage primer set for end-point (RT-PCR) and real-time (RT-qPCR) for CDV detection. Initially, we collected 194 complete/near-complete CDV genomes (GenBank) and analyzed them for highly conserved regions for primer design. We then assessed the in...
Source: Journal of Virological Methods - November 18, 2023 Category: Virology Authors: Alice Silveira Becker Tha ísa Regina Rocha Lopes Nat ália Hettwer Pedroso Jos é Valter Joaquim Silva Júnior Rudi Weiblen Eduardo Furtado Flores Source Type: research
Purification of Hepatitis C Virus Core Protein in Non-Denaturing Condition
In this study, we successfully solubilized bacterially expressed core protein using a high-salt and detergent-containing buffer and bypassed the denaturing-refolding process. Size-exclusion chromatography revealed three distinct peaks in the HCV-infected cell lysate, with the bacterially expressed soluble core protein corresponding to its second peak. Using a combination of affinity, size exclusion, and multi-modal chromatography purification techniques, we achieved a purity of >95% for the core protein. Analytical ultracentrifugation revealed monomer formation in the solution. Far UV Circular dichroism spectroscopy ide...
Source: Journal of Virological Methods - November 18, 2023 Category: Virology Authors: Kyo Izumida Yumiko Hara Yukio Furukawa Kotaro Ishida Keisuke Tabata Eiji Morita Source Type: research
Novel high-coverage primers for detection of canine morbillivirus by end-point and real-time RT-PCR assays
J Virol Methods. 2023 Nov 16:114853. doi: 10.1016/j.jviromet.2023.114853. Online ahead of print.ABSTRACTCanine distemper virus (CDV) is a major threat to domestic dogs and wildlife worldwide. Molecular assays are the most sensitive and specific tests to diagnose the disease, however, the high CDV genetic variability may compromise laboratory diagnosis. Herein, we designed a high-coverage primer set for end-point (RT-PCR) and real-time (RT-qPCR) for CDV detection. Initially, we collected 194 complete/near-complete CDV genomes (GenBank) and analyzed them for highly conserved regions for primer design. We then assessed the in...
Source: Journal of Virological Methods - November 18, 2023 Category: Virology Authors: Alice Silveira Becker Tha ísa Regina Rocha Lopes Nat ália Hettwer Pedroso Jos é Valter Joaquim Silva Júnior Rudi Weiblen Eduardo Furtado Flores Source Type: research
Purification of Hepatitis C Virus Core Protein in Non-Denaturing Condition
In this study, we successfully solubilized bacterially expressed core protein using a high-salt and detergent-containing buffer and bypassed the denaturing-refolding process. Size-exclusion chromatography revealed three distinct peaks in the HCV-infected cell lysate, with the bacterially expressed soluble core protein corresponding to its second peak. Using a combination of affinity, size exclusion, and multi-modal chromatography purification techniques, we achieved a purity of >95% for the core protein. Analytical ultracentrifugation revealed monomer formation in the solution. Far UV Circular dichroism spectroscopy ide...
Source: Journal of Virological Methods - November 18, 2023 Category: Virology Authors: Kyo Izumida Yumiko Hara Yukio Furukawa Kotaro Ishida Keisuke Tabata Eiji Morita Source Type: research
