Establishment of Swine Primary Nasal, Tracheal, and Bronchial Epithelial Cell Culture Models for the Study of Influenza Virus Infection
J Virol Methods. 2024 Apr 26:114943. doi: 10.1016/j.jviromet.2024.114943. Online ahead of print.ABSTRACTWe established primary porcine nasal, tracheal, and bronchial epithelial cells that recapitulate the physical and functional properties of the respiratory tract and have the ability to fully differentiate. Trans-well cultures demonstrated increased transepithelial electrical resistance over time the presence of tight junctions as demonstrated by immunohistochemistry. The nasal, tracheal, and bronchial epithelial cells developed cilia, secreted mucus, and expressed sialic acids on surface glycoproteins, the latter which a...
Source: Journal of Virological Methods - April 28, 2024 Category: Virology Authors: Madelyn Krunkosky Thomas M Krunkosky Victoria Meliopoulos Constantinos S Kyriakis Stacey Schultz-Cherry S Mark Tompkins Source Type: research
Establishment of Swine Primary Nasal, Tracheal, and Bronchial Epithelial Cell Culture Models for the Study of Influenza Virus Infection
J Virol Methods. 2024 Apr 26:114943. doi: 10.1016/j.jviromet.2024.114943. Online ahead of print.ABSTRACTWe established primary porcine nasal, tracheal, and bronchial epithelial cells that recapitulate the physical and functional properties of the respiratory tract and have the ability to fully differentiate. Trans-well cultures demonstrated increased transepithelial electrical resistance over time the presence of tight junctions as demonstrated by immunohistochemistry. The nasal, tracheal, and bronchial epithelial cells developed cilia, secreted mucus, and expressed sialic acids on surface glycoproteins, the latter which a...
Source: Journal of Virological Methods - April 28, 2024 Category: Virology Authors: Madelyn Krunkosky Thomas M Krunkosky Victoria Meliopoulos Constantinos S Kyriakis Stacey Schultz-Cherry S Mark Tompkins Source Type: research
Establishment of Swine Primary Nasal, Tracheal, and Bronchial Epithelial Cell Culture Models for the Study of Influenza Virus Infection
J Virol Methods. 2024 Apr 26:114943. doi: 10.1016/j.jviromet.2024.114943. Online ahead of print.ABSTRACTWe established primary porcine nasal, tracheal, and bronchial epithelial cells that recapitulate the physical and functional properties of the respiratory tract and have the ability to fully differentiate. Trans-well cultures demonstrated increased transepithelial electrical resistance over time the presence of tight junctions as demonstrated by immunohistochemistry. The nasal, tracheal, and bronchial epithelial cells developed cilia, secreted mucus, and expressed sialic acids on surface glycoproteins, the latter which a...
Source: Journal of Virological Methods - April 28, 2024 Category: Virology Authors: Madelyn Krunkosky Thomas M Krunkosky Victoria Meliopoulos Constantinos S Kyriakis Stacey Schultz-Cherry S Mark Tompkins Source Type: research
Establishment of Swine Primary Nasal, Tracheal, and Bronchial Epithelial Cell Culture Models for the Study of Influenza Virus Infection
J Virol Methods. 2024 Apr 26:114943. doi: 10.1016/j.jviromet.2024.114943. Online ahead of print.ABSTRACTWe established primary porcine nasal, tracheal, and bronchial epithelial cells that recapitulate the physical and functional properties of the respiratory tract and have the ability to fully differentiate. Trans-well cultures demonstrated increased transepithelial electrical resistance over time the presence of tight junctions as demonstrated by immunohistochemistry. The nasal, tracheal, and bronchial epithelial cells developed cilia, secreted mucus, and expressed sialic acids on surface glycoproteins, the latter which a...
Source: Journal of Virological Methods - April 28, 2024 Category: Virology Authors: Madelyn Krunkosky Thomas M Krunkosky Victoria Meliopoulos Constantinos S Kyriakis Stacey Schultz-Cherry S Mark Tompkins Source Type: research
Establishment of Swine Primary Nasal, Tracheal, and Bronchial Epithelial Cell Culture Models for the Study of Influenza Virus Infection
J Virol Methods. 2024 Apr 26:114943. doi: 10.1016/j.jviromet.2024.114943. Online ahead of print.ABSTRACTWe established primary porcine nasal, tracheal, and bronchial epithelial cells that recapitulate the physical and functional properties of the respiratory tract and have the ability to fully differentiate. Trans-well cultures demonstrated increased transepithelial electrical resistance over time the presence of tight junctions as demonstrated by immunohistochemistry. The nasal, tracheal, and bronchial epithelial cells developed cilia, secreted mucus, and expressed sialic acids on surface glycoproteins, the latter which a...
Source: Journal of Virological Methods - April 28, 2024 Category: Virology Authors: Madelyn Krunkosky Thomas M Krunkosky Victoria Meliopoulos Constantinos S Kyriakis Stacey Schultz-Cherry S Mark Tompkins Source Type: research
Advancing broad bean true mosaic virus detection using conventional RT-PCR and real-time RT-PCR with novel primer set design
J Virol Methods. 2024 Apr 25:114946. doi: 10.1016/j.jviromet.2024.114946. Online ahead of print.ABSTRACTBroad bean true mosaic virus (BBTMV) infects broad beans and peas, reducing yield. As BBTMV is transmitted through broad beans, many countries have implemented regulations to prevent the distribution of infected seeds. Currently, enzyme-linked immunosorbent assay (ELISA) is commonly used to detect BBTMV. While the PCR-based method is preferred for seed virus detection due to its sensitivity and speed. A BBTMV-specific PCR detection method has not yet been reported. A universal detection method currently exists that utili...
Source: Journal of Virological Methods - April 27, 2024 Category: Virology Authors: Fumino Nito Hitoshi Oya Takayuki Matsuura Hironobu Yanagisawa Source Type: research
Development and evaluation of a multiplex real-time RT-PCR assay for simultaneous detection of H5, H7, and H9 subtype avian influenza viruses
In this study, the multiplex H5/7/9 rRT-PCR assay was developed with sets of primers and probe updated or newly designed to simultaneously detect the H5, H7 and H9 genes. Multiplex H5/7/9 rRT-PCR showed 100% specificity without cross-reactivity with other subtypes of AIVs and avian disease-causing viruses or bacteria, and the limit of detection was 1 to 10 EID50/0.1ml (50% egg infectious dose). Artificial mixed infections with the three different subtypes could be detected accurately with high analytical sensitivity even under highly biased relative molecular ratios by balancing the reactivities of each subtype by modifyin...
Source: Journal of Virological Methods - April 26, 2024 Category: Virology Authors: Se-Hee An Na-Yeong Kim Gyeong-Beom Heo Yong-Myung Kang Youn-Jeong Lee Kwang-Nyeong Lee Source Type: research
Development and evaluation of a multiplex real-time RT-PCR assay for simultaneous detection of H5, H7, and H9 subtype avian influenza viruses
In this study, the multiplex H5/7/9 rRT-PCR assay was developed with sets of primers and probe updated or newly designed to simultaneously detect the H5, H7 and H9 genes. Multiplex H5/7/9 rRT-PCR showed 100% specificity without cross-reactivity with other subtypes of AIVs and avian disease-causing viruses or bacteria, and the limit of detection was 1 to 10 EID50/0.1ml (50% egg infectious dose). Artificial mixed infections with the three different subtypes could be detected accurately with high analytical sensitivity even under highly biased relative molecular ratios by balancing the reactivities of each subtype by modifyin...
Source: Journal of Virological Methods - April 26, 2024 Category: Virology Authors: Se-Hee An Na-Yeong Kim Gyeong-Beom Heo Yong-Myung Kang Youn-Jeong Lee Kwang-Nyeong Lee Source Type: research
Heparin-mediated PCR interference in SARS-CoV-2 assays and subsequent reversal with heparinase I
This study aimed to investigate the magnitude of heparin interference across various clinical laboratory testing platforms, and the reversal of any interference by degradation of heparin using the heparinase I enzyme in nasopharyngeal swab (NP) samples for SARS-CoV-2 analysis by RT-qPCR. Heparin-mediated PCR interference was evident at heparin concentrations as low as 10 IU/mL across all platforms tested, with the exclusion of the Hologic Panther Aptima SARS-CoV-2 assay. Rates of false negative or invalid results and falsely elevated cycle threshold (Ct) values increased with increasing heparin concentrations on all platfo...
Source: Journal of Virological Methods - April 22, 2024 Category: Virology Authors: K Edwards T Corocher Y Hersusianto D Campbell K Subbarao J A Neil P Monagle P Ho Source Type: research
Validation of a severe acute respiratory syndrome coronavirus 2 microneutralization assay for evaluation of vaccine immunogenicity
J Virol Methods. 2024 Apr 20:114945. doi: 10.1016/j.jviromet.2024.114945. Online ahead of print.ABSTRACTAs variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to emerge, assessment of vaccine immunogenicity remains a critical factor to support continued vaccination. To this end, an in vitro microneutralization (MN50) assay was validated to quantitate SARS-CoV-2 neutralizing antibodies against prototype and variant strains (Beta, Delta, Omicron BA.1, Omicron BA.5, and XBB.1.5) in human serum. For the prototype strain, the MN50 assay met acceptance criteria for inter-/intra-assay precision, spec...
Source: Journal of Virological Methods - April 22, 2024 Category: Virology Authors: Stephanie Hamilton Mingzhu Zhu Shane Cloney Clark Penny Mayes Jen Fenner Leah Cui Rongman Cai Raj Kalkeri Louis F Fries Melinda Pryor Joyce Plested Source Type: research
Heparin-mediated PCR interference in SARS-CoV-2 assays and subsequent reversal with heparinase I
This study aimed to investigate the magnitude of heparin interference across various clinical laboratory testing platforms, and the reversal of any interference by degradation of heparin using the heparinase I enzyme in nasopharyngeal swab (NP) samples for SARS-CoV-2 analysis by RT-qPCR. Heparin-mediated PCR interference was evident at heparin concentrations as low as 10 IU/mL across all platforms tested, with the exclusion of the Hologic Panther Aptima SARS-CoV-2 assay. Rates of false negative or invalid results and falsely elevated cycle threshold (Ct) values increased with increasing heparin concentrations on all platfo...
Source: Journal of Virological Methods - April 22, 2024 Category: Virology Authors: K Edwards T Corocher Y Hersusianto D Campbell K Subbarao J A Neil P Monagle P Ho Source Type: research
Validation of a severe acute respiratory syndrome coronavirus 2 microneutralization assay for evaluation of vaccine immunogenicity
J Virol Methods. 2024 Apr 20:114945. doi: 10.1016/j.jviromet.2024.114945. Online ahead of print.ABSTRACTAs variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to emerge, assessment of vaccine immunogenicity remains a critical factor to support continued vaccination. To this end, an in vitro microneutralization (MN50) assay was validated to quantitate SARS-CoV-2 neutralizing antibodies against prototype and variant strains (Beta, Delta, Omicron BA.1, Omicron BA.5, and XBB.1.5) in human serum. For the prototype strain, the MN50 assay met acceptance criteria for inter-/intra-assay precision, spec...
Source: Journal of Virological Methods - April 22, 2024 Category: Virology Authors: Stephanie Hamilton Mingzhu Zhu Shane Cloney Clark Penny Mayes Jen Fenner Leah Cui Rongman Cai Raj Kalkeri Louis F Fries Melinda Pryor Joyce Plested Source Type: research
Heparin-mediated PCR interference in SARS-CoV-2 assays and subsequent reversal with heparinase I
This study aimed to investigate the magnitude of heparin interference across various clinical laboratory testing platforms, and the reversal of any interference by degradation of heparin using the heparinase I enzyme in nasopharyngeal swab (NP) samples for SARS-CoV-2 analysis by RT-qPCR. Heparin-mediated PCR interference was evident at heparin concentrations as low as 10 IU/mL across all platforms tested, with the exclusion of the Hologic Panther Aptima SARS-CoV-2 assay. Rates of false negative or invalid results and falsely elevated cycle threshold (Ct) values increased with increasing heparin concentrations on all platfo...
Source: Journal of Virological Methods - April 22, 2024 Category: Virology Authors: K Edwards T Corocher Y Hersusianto D Campbell K Subbarao J A Neil P Monagle P Ho Source Type: research
Validation of a severe acute respiratory syndrome coronavirus 2 microneutralization assay for evaluation of vaccine immunogenicity
J Virol Methods. 2024 Apr 20:114945. doi: 10.1016/j.jviromet.2024.114945. Online ahead of print.ABSTRACTAs variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to emerge, assessment of vaccine immunogenicity remains a critical factor to support continued vaccination. To this end, an in vitro microneutralization (MN50) assay was validated to quantitate SARS-CoV-2 neutralizing antibodies against prototype and variant strains (Beta, Delta, Omicron BA.1, Omicron BA.5, and XBB.1.5) in human serum. For the prototype strain, the MN50 assay met acceptance criteria for inter-/intra-assay precision, spec...
Source: Journal of Virological Methods - April 22, 2024 Category: Virology Authors: Stephanie Hamilton Mingzhu Zhu Shane Cloney Clark Penny Mayes Jen Fenner Leah Cui Rongman Cai Raj Kalkeri Louis F Fries Melinda Pryor Joyce Plested Source Type: research
Real-world performance of the NeuMoDx ™ HCV Quant Test for quantification of hepatitis C virus (HCV)-RNA
J Virol Methods. 2024 Apr 11;327:114937. doi: 10.1016/j.jviromet.2024.114937. Online ahead of print.ABSTRACTQuantification of hepatitis C virus (HCV)-RNA in serum or plasma samples is an essential parameter in HCV diagnostics. Here, the NeuMoDx™Molecular System (Qiagen) was tested for the most common HCV genotypes and compared to the cobas c6800 system (Roche). HCV-RNA from 131 plasma/serum samples from chronically infected patients was determined in parallel on the NeuMoDx and c6800 systems. Linearity was analysed using the four most common HCV genotypes (1-4) in our cohort. The coefficient of variation (CV) within (int...
Source: Journal of Virological Methods - April 13, 2024 Category: Virology Authors: Nadine L übke Andreas Walker Martin Obermeier Jennifer Camdereli Martha Paluschinski Lara Walotka Anna-Kathrin Schupp Inga Tometten Sandra Hauka Eva Heger J örg Timm Source Type: research
