Last data update: Apr 29, 2024. (Total: 46658 publications since 2009)
Records 1-5 (of 5 Records) |
Query Trace: Lin SC[original query] |
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Development and Evaluation of a TaqMan Real-Time PCR Assay for the Rapid Detection of Cross-Contamination of RD (Human) and L20B (Mouse) Cell Lines Used in Poliovirus Surveillance.
Ahmad A , Lee JR , Metz JM , Tang X , Lin SC , Bagarozzi DAJr , Petway D , Herzegh O . J Virol Methods 2021 300 114354 BACKGROUND: The cross-contamination of cell lines in culture is a persistent problem. Genetically modified L20B (Mouse) and RD (Human Rhabdomyosarcoma) cell lines are commonly used in poliovirus research, surveillance, and diagnostics. Cross-contamination between these cell lines leads to unreproducible results and unreliable surveillance data, negatively affecting public health. The gold standard method for cell authentication is Short Tandem Repeats analysis, which is time-consuming and expensive. The disadvantage of STR is limited detection of interspecies contamination. METHODS: This assay targets the mitochondrial cytochrome c oxidase subunit I (MTCO1) gene, a highly conserved and emergent DNA barcode region for detection of cross-contamination in RD and L20B cell lines. The MagNA Pure Compact instrument and ABI 7500 Fast Dx Real-time PCR systems were used for DNA extraction and to perform real-time PCR respectively. RESULTS: The newly developed assay is very sensitive with a limit of detection of 100 RD cells/1 million L20B/mL. The amplification efficiency and R(2)-value were 102.26% and 0.9969 respectively. We evaluated specificity of the assay with five human and four mouse cell lines, as well as monkey and rat cell lines. The assay showed no cross-reactivity with genomic DNA from human, mouse, rat, or monkey cell lines. The analytical sensitivity was also evaluated by spiking varying amounts of RD cells (0.001% - 10%) into L20B cells. There was no difference in C(T) values when running single-plex or duplex PCR reactions with similar experimental conditions. CONCLUSIONS: We have developed and validated a TaqMan real-time PCR assay, a sensitive method for the detection of cross-contamination of RD and L20B cell lines. |
Improved specificity and reduced subtype cross-reactivity for antibody detection by ELISA using globular head domain recombinant hemagglutinin
Li ZN , Carney PJ , Lin SC , Li J , Chang JC , Veguilla V , Stevens J , Miller JD , Levine M , Katz JM , Hancock K . J Virol Methods 2014 209 121-5 The relative performance of ELISA using globular head domain (GH) and ectodomain hemagglutinins (HAs) as antigens to detect influenza A virus IgG antibody responses was assessed. Assay sensitivity and subtype cross-reactivity were evaluated using sera collected from recipients of monovalent H5N1 vaccine and A(H1N1)pdm09 virus-infected persons. Assay specificity was determined using collections of sera from either individuals unexposed to either H5N1 or A(H1N1)pdm09 viruses or exposed to H5N1 or A(H1N1)pdm09 viruses through vaccination or infection, respectively. ELISA using GH HA showed a similar degree of sensitivity, significantly higher specificity, and significantly lower subtype cross-reactivity compared to ELISA using ectodomain HA. |
IgM, IgG, and IgA antibody responses to influenza A(H1N1)pdm09 hemagglutinin in infected persons during the first wave of the 2009 pandemic in the United States
Li ZN , Lin SC , Carney PJ , Li J , Liu F , Lu X , Liu M , Stevens J , Levine M , Katz JM , Hancock K . Clin Vaccine Immunol 2014 21 (8) 1054-60 Novel influenza A(H1N1)pdm09 virus caused an influenza pandemic in 2009. IgM, IgG, and IgA antibody responses to A(H1N1)pdm09 hemagglutinin (HA) following A(H1N1)pdm09 virus infection were analyzed to understand antibody isotype responses. Age-matched control sera collected from U.S. residents in 2007 and 2008 were used to establish baseline levels of cross-reactive antibodies. IgM responses often used as an indicator of primary virus infection were mainly detected in young patient groups (≤5 yrs and 6-15 yrs), not in older age group, despite the genetic and antigenic differences between the HA of A(H1N1)pdm09 virus and pre-2009 seasonal H1N1 viruses. IgG and IgA responses to A(H1N1)pdm09 HA were detected in all age groups of infected persons. In persons aged 17-80 years, paired acute and convalescent serum samples demonstrated a four-fold or greater increase in the IgG and IgA responses to A(H1N1)pdm09 HA in 80% and 67% of A(H1N1)pdm09 virus-infected persons, respectively. The IgG antibody response to A(H1N1)pdm09 HA was cross-reactive with HAs from H1, H3, H5, and H13 subtypes suggesting that infections with subtypes other than A(H1N1)pdm09 could result in false positives by ELISA. Lower sensitivity compared to hemagglutination inhibition and microneutralization assays and the detection of cross-reactive antibodies against homologous and heterologous subtype are major drawback for application of ELISA in influenza serologic studies. |
Presence of antibodies against genogroup VI norovirus in humans
Mesquita JR , Costantini VP , Cannon JL , Lin SC , Nascimento MS , Vinje J . Virol J 2013 10 176 BACKGROUND: Noroviruses are important enteric pathogens in humans and animals. Recently, we reported a novel canine norovirus (CaNoV) in dogs with diarrhea belonging to a new genogroup (GVI). No data are available on exposure of humans to this virus. METHODS: Sera from 373 small animal veterinarians and 120 age-matched population controls were tested for IgG antibodies to CaNoV by a recombinant virus like particle based enzyme-linked immunosorbent assay. RESULTS: Antibodies to CaNoV were found in 22.3% of the veterinarians and 5.8% of the control group (p < 0.001). Mean corrected OD450 values for CaNoV antibodies were significantly higher in small animal veterinarians compared to the control group. CONCLUSIONS: These findings suggest that CaNoV may infect humans and small animal veterinarians are at an increased risk for exposure to this virus. Additional studies are needed to assess if this virus is able to cause disease in humans. |
Enhancing early identification and coordination of intervention services for young children with autism spectrum disorders: report from the Act Early Regional Summit Project
Peacock G , Lin SC . Disabil Health J 2012 5 (1) 55-9 BACKGROUND: Increasing prevalence of autism spectrum disorders (ASD) and the merits of early intervention support the importance of early identification and detection. The Act Early Initiative attempts to address the states' capacity to support this process of early identification and early intervention. OBJECTIVE: The Centers for Disease Control and Prevention (CDC) Health Resources and Services Administration (HRSA) collaborated with the Association of University Centers on Disabilities (AUCD) to develop strategies that will address state capacity for responding to the increasing demand for earlier identification, earlier diagnoses, and coordination of service systems for children with ASDs and other developmental disabilities (DD). METHODS: Act Early regional summits were held to engage stakeholders from the early detection and intervention community including parents, state agencies, provider groups, autism and related disability organizations, and academia. The stakeholders then used the Logic Model to facilitate the teams' planning process. The Logic Model enables teams to understand the strengths and gaps within their state resources and plan specific activities to achieve concrete outcomes. RESULTS: States identified opportunities and challenges in early identification of children with delay. One of the particular challenges identified were low income, rural and non-English speaking populations encountering more challenges than others in accessing diagnosis and early intervention services. CONCLUSIONS: The Summits are a unique model that demonstrates the importance of developing comprehensive state plans to advance the collaboration and coordination of early detection and intervention service systems for children with ASDs and related DDs from all racial, ethnic, geographical, and socioeconomic backgrounds. |
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