SARS-CoV-2 Nucleoprotein ELISA Kit (DEIASL017)

Regulatory status: For research use only, not for use in diagnostic procedures.

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Size
96T
Sample
Serum or plasma
Species Reactivity
Human
Intended Use
The SARS-CoV-2 N protein ELISA kit is to be used to detect and quantify protein levels of endogenous SARS-CoV-2 N protein.
Contents of Kit
1.Microplate-Antibody coated 96-well microplate (8 well × 12 strips), 1 plate
2.Protein standard-24000 pg/bottle, lyophilized, 2 bottles
3.Detection antibody, HRP-conjugated (100X)-120 μL/vial, 1 vial
4.Sample Diluent PT 4-30 mL/bottle, 1 bottle
5.Detection Diluent-30 mL/bottle, 1 bottle
6.Wash Buffer Concentrate (20X)-30 mL/bottle, 1 bottle
7.Tetramethylbenzidine Substrate (TMB)-12 mL/bottle, 1 bottle
8.Stop Solution-12 mL/bottle, 1 bottle
9.Plate Cover Seals, 3 pieces
Storage
Unopened Kit: Store at 2-8°C for 6 months or - 20°C for 12 months.
Opened Kit: All reagents stored at 2-8°C for 7 days. Please use a new standard for each assay.
Precision
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 20 times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 24 separate assays to assess inter-assay precision.
Detection Range
375-6000 pg/mL
Sensitivity
The minimum detectable dose of SARS-CoV-2 N protein is 38.0 pg/mL. This was determined by adding two standard deviations to the concentration corresponding to the mean O.D. of 20 zero standard replicates.
General Description
Coronaviruses are enveloped viruseswith a positive-sense RNA genome and with a nucleocapsid of helical symmetry. Coronavirus nucleoprotein slocalize to the cytoplasm and the nucleolus, a subnuclear structure, in both virus-infected primary cells and in cells transfected with plasmids that express N protein. Coronavirus N protein is required for coronavirus RNA synthesis and has RNA chaperone activity that may be involved in template switch. Nucleocapsid protein is a most abundant protein of coronavirus. During virion assembly, N protein bindsto viral RNA and leadsto formation of the helical nucleocapsid. Nucleocapsid protein is a highly immunogenic phosphoprotein also implicated in viral genome replication and in modulating cell signaling pathways. Because of the conservation of N protein sequence and its strong immunogenicity, the N protein of coronavirusis chosen as a diagnostic tool.
Standard Curve
These standard curves are provided for demonstration only. A standard curve should be generated for each set ofsamples assayed.

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References


Breastfed 13 month-old infant of a mother with COVID-19 pneumonia: a case report

INTERNATIONAL BREASTFEEDING JOURNAL

Authors: Yu, Yuanyuan; Li, Youjiang; Hu, Yingying; Li, Bin; Xu, Jian

Background In China, mothers with confirmed or suspected COVID-19 pneumonia are recommended to stop breastfeeding. However, the evidence to support this guidance is lacking. There have been relatively few cases reported about direct breastfeeding an infant by a mother with SARS-CoV-2 pneumonia. Therefore, it is necessary to assess the safety of breastfeeding and the possible protective effects of breast milk on infants. Case presentation This report analyzes the case of a mother who continued breastfeeding her 13 month-old child when both were diagnosed with confirmed COVID-19 pneumonia. We describe the clinical presentation, diagnosis, treatment, and outcome. The presence of SARS-CoV-2 nucleic acid was determined in maternal serum, breast milk, nasopharyngeal (NP) swabs and feces, and in infant serum, NP swabs and feces. IgM and IgG antibodies against SARS-CoV-2 were assessed in maternal serum and breast milk and in infant serum. SARS-CoV-2 nucleic acid was not detected in the breast milk, and antibodies against SARS-CoV-2 were detected in the mother's serum and milk. Conclusions The present case further confirms that the possibility of mother-to-child transmission about SARS-CoV-2 via breast milk alone was very small, and breast milk is safe for direct feeding of infants.

False-positive SARS-CoV-2 serology in 3 children with Kawasaki disease

DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE

Authors: To, Kelvin K. W.; Chua, Gilbert T.; Kwok, Ka Li; Wong, Joshua S. C.; Au, Dennis Chi Yu; Lam, Yuen Yu; Wong, Wilfred H. S.; Ho, Marco H. K.; Chan, Godfrey C. F.; Chui, Celine S. L.; Li, Xue; Tung, Keith T. S.; Wong, Rosa S.; Tso, Winnie W. Y.; Wong, Ian C. K.; Wong, Christina S. M.; Fong, Carol H. Y.; Chan, Kwok Hung; Yuen, Kwok Yung; Ip, Patrick; Kwan, Mike Y. W.

Background: Kawasaki disease (KD) is an acute febrile and eruptive disease with systemic vasculitis predominantly affecting young East Asian children. Recent reports showed that children with KD-like disease from KD low prevalence regions had positive SARS-CoV-2 serology despite a negative SARS-CoV-2 polymerase chain reaction (PCR) in respiratory samples. Objectives: To describe 3 pediatric Kawasaki Disease patients with false positive SARS-CoV-2 serology. Study design: We retrospectively recruited children with KD diagnosed during the COVID-19 outbreak in Hong Kong. Clinical characteristics and laboratory test results including SARS-CoV-2 PCR results were retrieved. We performed a microparticle-based immunoassay for the detection of IgG against nucleoprotein (NP) and spike protein receptor binding domain (RBD), and a microneutralization assay for the detection of neutralizing antibodies. Results: Three Chinese children with typical KD were identified. They had no epidemiological links with COVID-19 patients and tested negative for SARS-CoV-2 NPA PCR. Theywere treated with IVIG and aspirin, and were discharged without complications. Subsequently 2 of them were tested positive against anti-RBD and anti-NP antibodies and 1 was tested positive against anti- RBD antibodies. However, microneutralization assay showed that neutralizing antibodies were absent, suggesting a false-positive IgG result. Conclusion: Detection of neutralizing antibodies is recommended to confirm previous SARS-CoV-2 infection in IgGpositive but PCR-negative patients. (C) 2020 The Author(s). Published by Elsevier Inc.

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