scholarly journals Performance and usefulness of a novel automated immunoassay HISCL SARS-CoV-2 Antigen assay kit for the diagnosis of COVID-19

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kaori Saito ◽  
Tomohiko Ai ◽  
Akinori Kawai ◽  
Jun Matsui ◽  
Yoshiyuki Fukushima ◽  
...  
Keyword(s):  
Clinical Performance ◽  
Quantitative Polymerase Chain Reaction ◽  
Serum Samples ◽  
N Protein ◽  
Detection Systems ◽  
Copy Numbers ◽  
Antigen Assay ◽  
Polymerase Chain ◽  
Human Coronaviruses ◽  
Automated Immunoassay

AbstractHere, we aimed to evaluate the clinical performance of a novel automated immunoassay HISCL SARS-CoV-2 Antigen assay kit designed to detect the nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This kit comprises automated chemiluminescence detection systems. Western blot analysis confirmed that anti-SARS-CoV antibodies detected SARS-CoV-2N proteins. The best cut-off index was determined, and clinical performance was tested using 115 serum samples obtained from 46 patients with coronavirus disease 2019 (COVID-19) and 69 individuals who tested negative for COVID-19 through reverse transcription quantitative polymerase chain reaction (RT-qPCR). The HISCL Antigen assay kit showed a sensitivity of 95.4% and 16.6% in samples with copy numbers > 100 and < 99, respectively. The kit did not cross-react with human coronaviruses causing seasonal common cold and influenza, and none of the 69 individuals without COVID-19 were diagnosed with positive results. Importantly, 81.8% of the samples with low virus load (< 50 copy numbers) were diagnosed as negative. Thus, using HISCL antigen assay kits may reduce overdiagnosis compared with RT-qPCR tests. The rapid and high-throughput HISCL SARS-CoV-2 Antigen assay kit developed here proved suitable for screening infectious COVID-19 and may help control the pandemic.

scholarly journals Performance and Usefulness of a Novel Automated Immunoassay HISCL SARS-CoV-2 Antigen Assay Kit for the Diagnosis of COVID-19

2021 ◽  
Author(s):  
Kaori Saito ◽  
Tomohiko Ai ◽  
Akinori Kawai ◽  
Jun Matsui ◽  
Yoshiyuki Fukushima ◽  
...  
Keyword(s):  
Clinical Performance ◽  
Quantitative Polymerase Chain Reaction ◽  
Serum Samples ◽  
N Protein ◽  
Detection Systems ◽  
Copy Numbers ◽  
Antigen Assay ◽  
Polymerase Chain ◽  
Automated Immunoassay ◽  
Corona Viruses

Abstract Background: The aim of this study was to evaluate the clinical performance of a newly developed automated immunoassay HISCL SARS-CoV-2 Antigen assay kit designed to detect the nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Methods: The HISCL SARS-CoV-2 Antigen assay kit is composed of automated chemiluminescence detection systems. Western blot analysis confirmed that anti-SARS-CoV antibodies detected SARS-CoV-2 N proteins. The best cut-off index (COI) was determined using human serum samples obtained from coronavirus disease-2019 (COVID-19) patients and patients without COVID-19. To test the clinical performance, 115 samples obtained from 46 patients with COVID-19 and 69 individuals who tested negative for COD-19 using reverse transcription quantitative polymerase chain reaction (RT-qPCR) were used. Results: The HISCL antigen assay kit showed a sensitivity of 95.4 % in the samples with copy numbers of >100, and a sensitivity of 16.6 % in the samples with copy numbers of <99. The kit did not cross-react with other human corona viruses causing seasonal common cold (HCoV 229E, OC43, NL63, and HKU1) and influenza (H1N1, H3N2, and B), and none of the 69 individuals with negative RT-qPCR results were diagnosed as positive. Importantly, 81.8 % of the samples with low virus load (<50 copy numbers) were diagnosed as negative. Thus, use of the HISCL antigen assay kit may reduce overdiagnosis compared with RT-qPCR tests. Conclusion: The rapid and high-throughput HISCL SARS-CoV-2 Antigen assay kit developed in this study can be used as a suitable screening test for infectious COVID-19, and may play a role in controlling the pandemic.

scholarly journals No Evidence of Mosquito Involvement in the Transmission of Equine Hepacivirus (Flaviviridae) in an Epidemiological Survey of Austrian Horses

Viruses ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1014 ◽  
Author(s):  
Marcha Badenhorst ◽  
Phebe de Heus ◽  
Angelika Auer ◽  
Till Rümenapf ◽  
Birthe Tegtmeyer ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Single Case ◽  
Quantitative Polymerase Chain Reaction ◽  
Epidemiological Survey ◽  
Serum Samples ◽  
Prevalence Studies ◽  
The Family ◽  
Eastern Austria ◽  
Polymerase Chain ◽  
Hematophagous Arthropods

Prevalence studies have demonstrated a global distribution of equine hepacivirus (EqHV), a member of the family Flaviviridae. However, apart from a single case of vertical transmission, natural routes of EqHV transmission remain elusive. Many known flaviviruses are horizontally transmitted between hematophagous arthropods and vertebrate hosts. This study represents the first investigation of potential EqHV transmission by mosquitoes. More than 5000 mosquitoes were collected across Austria and analyzed for EqHV ribonucleic acid (RNA) by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Concurrently, 386 serum samples from horses in eastern Austria were analyzed for EqHV-specific antibodies by luciferase immunoprecipitation system (LIPS) and for EqHV RNA by RT-qPCR. Additionally, liver-specific biochemistry parameters were compared between EqHV RNA-positive horses and EqHV RNA-negative horses. Phylogenetic analysis was conducted in comparison to previously published sequences from various origins. No EqHV RNA was detected in mosquito pools. Serum samples yielded an EqHV antibody prevalence of 45.9% (177/386) and RNA prevalence of 4.15% (16/386). EqHV RNA-positive horses had significantly higher glutamate dehydrogenase (GLDH) levels (p = 0.013) than control horses. Phylogenetic analysis showed high similarity between nucleotide sequences of EqHV in Austrian horses and EqHV circulating in other regions. Despite frequently detected evidence of EqHV infection in Austrian horses, no viral RNA was found in mosquitoes. It is therefore unlikely that mosquitoes are vectors of this flavivirus.

scholarly journals Middle East respiratory syndrome coronavirus (MERS-CoV) in dromedary camels in Nigeria, 2015

2015 ◽  
Vol 20 (49) ◽  
Author(s):  
Daniel KW Chu ◽  
Jamiu O Oladipo ◽  
Ranawaka APM Perera ◽  
Sulaiman A Kuranga ◽  
Samuel MS Chan ◽  
...  
Keyword(s):  
Middle East ◽  
Arabian Peninsula ◽  
Phylogenetic Analyses ◽  
Quantitative Polymerase Chain Reaction ◽  
Middle East Respiratory Syndrome ◽  
Dromedary Camels ◽  
Chain Reaction ◽  
Serum Samples ◽  
Nasal Swabs ◽  
Polymerase Chain

Evidence of current and past Middle East respiratory syndrome coronavirus (MERS-CoV) infection in dromedary camels slaughtered at an abattoir in Kano, Nigeria in January 2015, was sought by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and serology. MERS-CoV RNA was detected in 14 (11%) of 132 nasal swabs and antibody in 126 (96%) of 131 serum samples. Phylogenetic analyses demonstrate that the viruses in Nigeria are genetically distinct from those reported in the Arabian peninsula.

DETECTION OF BAT CORONAVIRUS AND SPECIFIC ANTIBODIES IN CHESTNUT BAT (SCOTOPHILUS KUHLII) POPULATION IN CENTRAL TAIWAN

2016 ◽  
Vol 42 (01) ◽  
pp. 19-26 ◽  
Author(s):  
Bo-Gang Su ◽  
Hong Chang Chen ◽  
Hsi-Chi Cheng ◽  
Yi-Ning Chen
Keyword(s):  
Cross Reactivity ◽  
Rdrp Gene ◽  
Rt Pcr ◽  
Protein Fragment ◽  
Serum Samples ◽  
Protein Fragments ◽  
N Protein ◽  
Polymerase Chain ◽  
Central Taiwan ◽  
Bat Coronavirus

Bats can serve as natural reservoirs for severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome coronavirus (MERS-CoV). Investigating the prevalence of bat CoV is critical for assessing the risks of the outbreaks of emerging CoV. Chestnut bats (Scotophilus kuhlii) were captured in this study for detecting the partial RNA-dependent RNA polymerase (RdRp) gene in their feces through reverse transcription polymerase chain reaction (RT-PCR) and antibodies to the nucleocapsid (N) protein of bat CoV through western blotting (WB) analysis. Three recombinant N protein fragments (N1, N2, N3) of the isolated Scotophilus bat CoV/CYCU-S1/TW/2013 were expressed by Escherichia coli. WB analyses were performed with bat serum samples and the sera of a patient who recovered from a SARS-CoV infection. Fragment N2 contained a highly conserved motif among CoVs whereas N1 and N3 protein fragments were specific to the S. kuhlii bat CoV. A total of 32 fecal and 19 serum samples were collected in Changhua County and Yunlin County during 2013 and 2014. About 17 fecal samples tested positive for the RdRp gene with an overall prevalence of 53%. Sequences comparison showed that the Scotophilus bat CoV isolates in Taiwan belonged to the genus Alphacoronavirus and were closest to Scotophilus bat CoV/Hainan/China/2005 and Diliman1552G1/Philippines/2008, followed by porcine epidemic diarrhea coronavirus. Only one bat serum sample reacted positively to all 3[Formula: see text]N protein fragments. Cross-reactivity was observed between N2 protein fragment and the sera of a patient recovered from a SARS-CoV infection. The results indicated that Scotophilus bat CoV was circulating endemically in chestnut bat population in Taiwan.

scholarly journals Antibody Response against the SARS-CoV-2 Nucleocapsid Protein and Its Subdomains—Identification of Pre-Immunization Status by Human Coronaviruses with Multipanel Nucleocapsid Fragment Immunoblotting

COVID ◽  
2021 ◽  
Vol 1 (1) ◽  
pp. 105-114
Author(s):  
Sahra Pajenda ◽  
Sebastian Kapps ◽  
Thomas Reiter ◽  
Raimundo Freire ◽  
Veronique A. J. Smits ◽  
...  
Keyword(s):  
Nucleocapsid Protein ◽  
Posterior Part ◽  
Rapid Identification ◽  
Diagnostic Methods ◽  
Serum Samples ◽  
N Protein ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Global Pandemic ◽  
Human Coronaviruses

A novel beta coronavirus that emerged in late December 2019 triggered a global pandemic. Diagnostic methods for rapid identification of infected individuals were established in new biotechnological approaches. Vaccine production and application to individuals and measurement of SARS-CoV-2 antibodies also began. Serum samples from 240 health care workers were collected at three-month intervals over nine months. Indirect SARS-CoV-2 nucleocapsid IgG ELISA tests were used to identify humoral immune responses. All seropositive individuals and those with borderline ELISA values were tested with a specifically generated multipanel nucleocapsid fragment immunoblot. Of the 240 individuals, 24 showed seroconversion in ELISA after experiencing COVID-19. All of them showed a positive reaction against the full-length nucleocapsid protein in the immunoblot. The highest reactivity was seen either against fragment N(100–300) or in a minority against the posterior part N(200–419). In general, the staining pattern of COVID-19 patients showed four phenotypes. In contrast, three individuals classified as borderline by ELISA reacted exclusively with fragments N(1–220) and N(100–300) containing the octamer amino acid sequence FYYLGTGP, which is identical in human coronaviruses sharing this sequence with SARS-CoV-2. These represent a unique and thus fifth phenotype. This work suggests the existence of distinct phenotypic patterns of IgG production towards N-protein subdomains.

scholarly journals Multiple-Centre Clinical Evaluation of Rapid Recombinase-Aided Amplification Assays for Five Pathogens

2021 ◽  
Author(s):  
Xin-xin Shen ◽  
Dan-wen Nie ◽  
Hong Zhang ◽  
Zhi-fei Zhan ◽  
Yuan Gao ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Reverse Transcription ◽  
Clinical Performance ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Adenovirus ◽  
Molecular Diagnostic ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Polymerase Chain

Abstract Background: Recombinase-aided amplification(RAA) is a new, simple, and ultrafast isothermal molecular diagnostic technique performed within 30min at 39°C–42°C.In this study, we evaluated the clinical performance of four duplex RAA kits for hepatitis B virus(HBV), human adenovirus 3(HAdV3), human adenovirus 7(HAdV7), and Bordetella pertussis and one duplex reverse-transcription RAA (RT-RAA) kit for respiratory syncytial virus (RSV).Methods: A total of 392 sera and 374 respiratory tract samples were collected from five institutions in four China regions. Each RAA kit’s sensitivity and specificity were compared with those of real-time quantitative polymerase chain reaction(qPCR),real-time quantitative reverse-transcription polymerase chain reaction(qRT-PCR), or sequencing. Results: Compared with qPCR or qRT-PCR, the sensitivities of HBV RAA,RSV RT-RAA, and B.pertussis RAA were 97.55%,96.67%, and 100%,respectively,and all of the specificities were 100%.The total coincidence rates were 97.78%(383/392,95%CI:95.63%–98.85%),97.70%(212/217, 95%CI:94.57%–99.16%), and 100%(60/60,95%CI:92.80%–100%),respectively.The Kappa values were 0.977,0.947, and 1,respectively(P<0.05).Regarding the sequencing, the sensitivities of HAdV3 RAA and HAdV7 RAA were 100% and 97.37%, respectively,and all specificities were 100%.The total coincidence rates were 100%(97/97,95%CI:91.58%–100%) and 98.97%(96/97,95%CI:94.39%–99.82%),and the Kappa values were 1 and 0.978 (P<0.05),respectively.Conclusions: With comparable clinical performance, these RAA kits are suitable assays for rapidly detecting pathogens in resource-limited laboratories.

scholarly journals Exploring the Connection between Porphyromonas gingivalis and Neurodegenerative Diseases: A Pilot Quantitative Study on the Bacterium Abundance in Oral Cavity and the Amount of Antibodies in Serum

Biomolecules ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 845
Author(s):  
Raffaella Franciotti ◽  
Pamela Pignatelli ◽  
Claudia Carrarini ◽  
Federica Maria Romei ◽  
Martina Mastrippolito ◽  
...  
Keyword(s):  
Oral Cavity ◽  
Porphyromonas Gingivalis ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Serum Samples ◽  
Amyloid Peptides ◽  
Neurodegenerative Process ◽  
Oral Pathogens ◽  
Preliminary Study ◽  
Polymerase Chain

Recent studies support the hypothesis that microbes can seed some Alzheimer’s disease (AD) cases, leading to inflammation and overproduction of amyloid peptides. Porphyromonas gingivalis (Pg) is a keystone pathogen of chronic periodontitis and has been identified as risk factor for the development and progression of AD. The present preliminary study aimed to quantify Pg abundance in neurodegenerative disease (ND) patients compared with neurologic patients without neurodegenerative disorders (no-ND) and healthy controls (HC) to determine possible association between Pg abundance and neurodegenerative process. Pg was quantified on DNA extracted from the oral samples of 49 patients and 29 HC by quantitative polymerase chain reaction (qPCR). Anti-Pg antibodies were also detected on patient serum samples by enzyme-linked immunosorbent assays (ELISA). The Pg abundance in the oral cavity was significantly different among groups (p = 0.004). It was higher in ND than no-ND (p = 0.010) and HC (p = 0.008). The Pg abundance was correlated with the antibodies (p = 0.001) with different slopes between ND and no-ND (p = 0.037). Pg abundance was not correlated with oral indices and comorbidities. These results extend our understanding of the association between oral pathogens and AD to other neurodegenerative processes, confirming the hypothesis that oral pathogens can induce an antibody systemic response, influencing the progression of the disease.

scholarly journals MMP1 is a Promiseful Prognostic Biomarker and Correlating with Immune Infiltrates in Hepatocellular Carcinoma

2021 ◽  
Author(s):  
Lei Dai ◽  
Joseph Muggnyi ◽  
xingchen cai ◽  
shuqi mao ◽  
tongyue zhang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Reverse Transcription ◽  
Clinical Performance ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Adenovirus ◽  
Molecular Diagnostic ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Polymerase Chain

Abstract Background: Recombinase-aided amplification(RAA) is a new, simple, and ultrafast isothermal molecular diagnostic technique performed within 30min at 39°C–42°C.In this study, we evaluated the clinical performance of four duplex RAA kits for hepatitis B virus(HBV), human adenovirus 3(HAdV3), human adenovirus 7(HAdV7), and Bordetella pertussis and one duplex reverse-transcription RAA (RT-RAA) kit for respiratory syncytial virus (RSV).Methods: A total of 392 sera and 374 respiratory tract samples were collected from five institutions in four China regions. Each RAA kit’s sensitivity and specificity were compared with those of real-time quantitative polymerase chain reaction(qPCR),real-time quantitative reverse-transcription polymerase chain reaction(qRT-PCR), or sequencing. Results: Compared with qPCR or qRT-PCR, the sensitivities of HBV RAA,RSV RT-RAA, and B.pertussis RAA were 97.55%,96.67%, and 100%,respectively,and all of the specificities were 100%.The total coincidence rates were 97.78%(383/392,95%CI:95.63%–98.85%),97.70%(212/217, 95%CI:94.57%–99.16%), and 100%(60/60,95%CI:92.80%–100%),respectively. The Kappa values were 0.977,0.947, and 1,respectively(P<0.05).Regarding the sequencing, the sensitivities of HAdV3 RAA and HAdV7 RAA were 100% and 97.37%, respectively, and all specificities were 100%.The total coincidence rates were 100%(97/97,95%CI:91.58%–100%) and 98.97%(96/97,95%CI:94.39%–99.82%),and the Kappa values were 1 and 0.978 (P<0.05),respectively.Conclusions: With comparable clinical performance, these RAA kits are suitable assays for rapidly detecting pathogens in resource-limited laboratories.

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