Day: January 20, 2022

Expected use

The SARS-CoV-2/Influenza A&B/RSV RT-qPCR Combo Kit is CE marked, in vitro diagnostic real-time reverse transcriptase PCR (RT-PCR) assay intended to be used for the identification and quantification of COVID-19 viral RNA extracted from Nasopharyngeal swabs and/or oropharyngeal swabs. The assay is designed to be used with designated extraction systems and PCR platforms. The combined SARS-CoV-2/Influenza A&B/RSV RT-qPCR Kit (CE IVD) assay is intended for use by specifically instructed and trained qualified and trained clinical laboratory personnel in real-time PCR techniques and in vitro diagnostic procedures.

Purpose, Scope and Users

SARS-CoV-2 has initially named the 2019 novel coronavirus (2019-nCoV) and was identified by Chinese authorities after an outbreak of pneumonia in December 2019 in Wuhan, Hubei province, China. The WHO declared an international health emergency on January 31, 2020. The Centers for Disease Control and Prevention (CDC) and the World Health Organization (WHO) recommend random testing for the asymptomatic population and testing for anyone with symptoms of samples collected from the upper airways and/or lower airways specimens for the identification of SARS-CoV-2 and other respiratory viruses, such as influenza and RSV. RT-qPCR is the gold standard method to identify active infections.

The NIPD Genetics Combined SARS-CoV-2/Influenza A&B/RSV RT-qPCR Kit is a Multiplex real-time RT-qPCR assay for the direct qualitative detection of RNA from 4 viruses: the new coronavirus (SARS-CoV-2), influenza A, influenza B and respiratory A/B syncytial virus (RSV A/B) from human respiratory specimens collected through the Nasopharyngeal and/or oropharyngeal swab method. Validation was performed on the QIAquant 96 5plex real-time PCR instrument (230 V) Real-time PCR detection system. This test can distinguish between influenza A and B subtypes, but not differentiate between RSV A and B. RNA is isolated from respiratory samples, reverse transcribed and amplified using RT-qPCR and detected with four fluorescent dye probes, allowing multiplexing and co-detection by sample reaction.

Fundamental reason

Diagnosis of the aforementioned viruses can be challenging due to the likeness of the symptoms. However, discriminatory and early diagnosis is important for the disease management and treatment, therefore, real-time PCR assay can be a sensitive method and reliable method to detect SARS-CoV-2, influenza A/B and RSV viruses. NIPD Genetics is not responsible for the outcome of a patient’s treatment if the results are being used to guide the medical treatment of health professionals.

Quality control and validity of the results

SARS-CoV-2/Influenza A&B/RSV RT-qPCR Combo Kit Contains Positive Results (PTC) and negative template controls (NTC) to be able to control the PCR amplification. Both of them controls should be included in each run for proper quality control, interpretation of results, and for the results to be considered valid.

Introduction

• Coronaviruses

are single-stranded, positive-sense, non-segmented RNA viruses. They belong to the Coronaviridae family. They are the largest known RNA viruses with genomes ranging between 27 and 31.5 kb. There are six species of coronaviruses known to causes human diseases 229E, OC43, NL63, HKU1, MERS-CoV, and SARS-CoV. SARS-CoV-2, named as a 2019 novel coronavirus (2019-nCoV) is responsible for the current pandemic that affects millions of people around the world. The virus is transmitted from human to human through respiratory droplets by direct or indirect contact.

Symptoms of SARS-CoV-2 can appear 2 to 14 days after exposure to the virus and at most common symptoms are fever, dry cough, fatigue. Less common symptoms include aches and pains, nasal congestion, sore throat, diarrhoea, loss of smell (anosmia) or loss of taste (ageusia), etc. The strongest and most consistent evidence of increased risk comes from patients with a BMI greater than 30 kg/m2, type 2 diabetes mellitus and with underlying lung and heart conditions, who are more likely to develop complications and require ICU admission and oxygen support.

• Influenza viruses

are negative-sense viruses that belong to the Orthomyxoviridae family. The most common influenza viruses that infect humans are influenza A and B. The most common symptoms include fever, cough, sore throat, nasal congestion, and discharge that can lead to more serious complications of pneumonia. age and underlying pulmonary medical conditions (eg, COPD, IPF, asthma, smokers) increase the risk of developing complications.

• Human respiratory syncytial viruses (RSV)

are non-segmented, negative, viruses with single-stranded linear RNA genome belonging to the Paramyxoviridae family. RSV causes respiratory infections in humans, including bronchitis, pneumonia, and chronic obstructive pulmonary infections. Common symptoms include runny nose discharge, low-grade fever, cough, sore throat, headache, and wheezing. All of the aforementioned viruses are transmitted between humans in three ways: (1) directly contact with infected people; (2) by contact with contaminated objects (such as toys, doorknobs); and (3) by inhalation of virus-laden aerosols.

General rules

1. Transport and storage conditions

Reagents are shipped and stored at -20°C until the expiration date, as indicated on the label. To minimize freeze-thaw cycles and preserve the integrity of the PTC, we recommend aliquoting and storing at -20°C. All components must be kept away from sunlight.

2. Training requirements

Testing for the presence of viral RNA should be performed in a laboratory equipped with trained personnel to carry out the pertinent technical procedures according to the Laboratory standards of the Occupational Safety and Health Administration (OSHA). Refer to the World Health Organization Interim Guidance on Laboratory Biosafety and the Centers for Disease Control and Prevention (CDC) Guidelines for Interim Laboratory Biosafety guidelines for the handling and processing of samples associated with SARS-CoV-2.

3. Precautions and Recommendations

• The procedures in this manual must be followed as described. any deviation may result in assay failure or cause erroneous results and interpretation.
• GLP is required to ensure kit performance, with care taken to avoid contamination of kit components. Components believed to have been converted Contaminated should be disposed of as standard laboratory waste in a sealed container ziplock bag or plastic bag.
• Sample samples must be collected, transported, and stored in accordance with appropriate laboratory guidelines. For more information, see the CDC3 guidelines for “Interim Guidelines for the Collection, Handling, and Analysis of Clinical Specimens for COVID-19″.
• Specimens must be processed within 4 hours of collection. Samples that will be processed after the 4-hour post-harvest window should be stored between 2-8°C for up to 72 hours.
• Thaw reagents at room temperature before use and keep on ice.
• Shelf life of reagents is 12 months when properly stored.
• Do not use reagents past the expiration date. After the expiration date, the quality of the warranty is no longer valid.
• Do not mix reagents from different kits and/or lots and/or from another supplier.
• Wear personal protective equipment (PPE), such as disposable gloves, goggles, and mask during sample collection and processing.
• Handle all specimens as if they were infectious using GLP and Safety Standards and Occupational Health Administration (OSHA) Laboratory Standard (29 CFR 1910.1450).
• Carry out all manipulations of possible live virus samples within a class II (or higher) biological safety cabinet.
• Dispose of waste in accordance with local, state and federal regulations.
• Regular decontamination of commonly used equipment is recommended, especially micropipettes and work surfaces with at least 70% (v/v) ethanol. Yes is working with RNA, to avoid degradation, it is recommended for the processing area banks to clean with RNase AWAY ® or 10% freshly prepared Bleach.
• Consult the safety data sheet (SDS) before using this kit, which is available on request.
• Please refer to the reference manual of each real-time qPCR instrument for additional information, warnings, precautions, procedures and data analysis.

The Myth of IgG Food Panel Testing

IgGEs Food Panel Test Very common for patients to feel they have food allergies or food intolerances/sensitivities. These terms or labels are often used interchangeably. However, it is important to understand that allergies are very different from intolerances or sensitivities. There are excellent materials on this website to help you distinguish between the two. With a food allergy, the body mounts an immune response to the food, and this can be dangerous. With an intolerance or sensitivity, the body may simply not be processing or digesting food properly and this is not really dangerous (although it can obviously be uncomfortable).

More classic food intolerances (such as lactose intolerance) cause patients to experience bloating, fullness, stomach pain, gas, and/or diarrhoea when they eat too much food. This is because the body is not digesting food properly, leading to a buildup of air and gases in the stomach and intestines. Other patients experience headaches, fatigue, “brain fog” or stomach pain with various foods or additives/preservatives. Oftentimes, patients feel that multiple foods may be causing these symptoms and are hoping to find a single test that will tell them exactly which foods to avoid so they can simply feel better.

Unfortunately, there is no single test that can give you this answer. One test that claims to be able to diagnose food sensitivities and is commonly available in the food IgG test. This test, offered by several companies, reports IgG levels for multiple foods (typically 90 to 100 foods with a single panel test) and claims that eliminating foods with high IgG levels can lead to improvement in multiple symptoms. Some websites even report that diets using this test can help with symptoms of irritable bowel syndrome, autism, cystic fibrosis, rheumatoid arthritis, and epilepsy.

It is important to understand that this test has never been scientifically proven to do what it claims to do. The scientific studies provided to support the use of this test are often out of date, in unaccredited journals, and many have not even used the IgG test in question. The presence of IgG is probably a normal response of the immune system to food exposure. In fact, higher levels of IgG4 to foods may simply be associated with tolerance to those foods.

COVID-19 Ag Rapid Test

The COVID-19 Rapid Antigen Test, a lateral flow chromatographic immunoassay, looks for protein fragments of the virus and thus its direct physical presence in the body. It is performed using a swab from the oral cavity or from the nasopharynx. This is extracted into a buffer solution and then pipetted into the test cassette. A qualitative result is ready in less than 20 minutes.

The diagnostic sensitivity is 97.56% (ct value: 20 – 30) and the diagnostic specificity is >99.9%. That means positive antigen test results are highly accurate. However, as there is always a residual risk of a false negative result if, in doubt, a PCR test should be performed to confirm negative results. The choice between antibody, antigen, and PCR testing is not a one or other decision. Rather, the best tactic is a combined application.

Test properties:

• Optimized testing process
• High specificity (>99.9%)
• Highly sensitive with high viral loads: 97.56% (ct value: 20 – 30)
• Easy to use
• Fast and reliable test results in just 15 minutes
• Testing can be performed using nasal, nasopharyngeal, and oropharyngeal specimens.
• It can be stored at room temperature.
• All test components, including sterile swabs, are included.
• No cross-reactivity with seasonal coronaviruses (such as hCoV-229E, -HKU1, -NL63, and -OC43) or influenza viruses (such as influenza A/B)

Importance of the antigen test

While the COVID-19 IgG/IgM Rapid Test detects the virus indirectly, the COVID-19 Rapid Antigen Test detects it directly by checking for the presence of specific proteins of the SARS-CoV-2 virus. Therefore, antigen detection closes a diagnostic gap. The nucleocapsid (N) protein is ideal for virus detection because it is highly abundant in the virus and specific enough for SARS-CoV-2. It is one of the 4 most important structural proteins of the virus and is involved in the replication, transcription and packaging of the viral genome.

The COVID-19 Rapid Antigen Test is a lateral flow chromatographic immunoassay for the qualitative detection of SARS-CoV-2 viral nucleoprotein antigens in human nasal- and oropharyngeal specimens. The test is intended to be used as an aid in the diagnosis of SARS-CoV-2 infections. It should be noted that the concentration of viral nucleoprotein antigens may vary during the course of the disease and may fall below the detection limit of the test. The possible infectiousness of test subjects cannot be excluded based on negative test results.

Description

This product uses “real-time fluorescent quantitative PCR technology” combined with self-developed “real-time dynamic precision temperature control technology”, which can be used in various fluorescent quantitative PCR detection projects to achieve accurate detection. rapid nucleic acid With powerful features and flexible operation, DNA and RNA PCR amplification and detection experiments can be completed in a compact and portable environment.

Using a special reverse anti-pollution open cap design, can effectively prevent contamination caused by accidental bursting of the hot cap, simplify the experimental operation process, and combine with efficient and fast PCR reagents to provide a complete solution for SARS. -CoV- 2 detection and other projects.

Additional Information

1. Equipment Features

• ORF1ab and N gene
• RNase P as an internal control
• CT cutoff: ≤30
• LoD: 500 copies/mL
• Throat smear or sputum
• This kit can also be compatible with ABI7500, BIORAD CFX96, AGS4800

2. Operation Steps (Combine with Nucleic Acid Releasing Reagent DA0940)

• Sampling and inactivation
• Rock and rest for 5 min, then centrifuge
• Add the extracted sample to the PCR tubes with PCR reagent
• Perform PCR, within 0.5 hours to obtain the result

Security

The following basic safety measures should be observed during the operation, maintenance and repair of the instrument. If the notes, warnings or cautions specified in the manual are neglected, the basic protection provided by the instrument may be affected. Meanwhile, it may also damage the security level of the design and manufacture of instruments and the intended scope of application.

It is not allowed to change the parts of the instrument without permission, and the inspection and maintenance of the instrument must be completed by engineers licensed by Hangzhou AnYu Technologies Co., Ltd. If the equipment is not used in the manner specified in this manual, the protection provided by the equipment may be damaged

Instrument maintenance

The wells and hot cap of the instrument should be periodically cleaned with special brushes bathed in a little absolute alcohol to guarantee complete contact between tubes and wells and high thermal conductivity while free of pollution. Use a soft cloth to do daily maintenance to clean the surface and wells. If there is any stain on the surface, you can clean it with a dust-free cloth with 75% alcohol. Do not use chlorine disinfectants, moisturizers, antibacterials soaps, aromatic solutions, propanol or isopropanol to clean the equipment, as they are types of cleaning or sanitizing agent is easy to have a chemical reaction with parts of the equipment or materials contained in the equipment and cause danger.

Perform only the maintenance mentioned in this manual. Before using any cleaning and disinfection method not described in this document, please contact the technical support personnel of the Company or authorized agency to ensure that the method used will not damage the instrument. If you have any questions on the compatibility of disinfectants or cleaning agents with pieces of equipment or materials contained in the equipment, consult the Company or an authorized agency.

Expected use

• In vitro diagnostic medical devices: IVD
• Diagnosis of SARS-CoV-2 infection
• Respiratory samples such as oropharyngeal or nasopharyngeal swabs

Features and Benefits

• One-step RT-PCR in a single tube
• Since the human RNase P gene is used as an internal control, it is possible to check whether the test result is correct, accurate or not by monitoring the whole testing process such as sample collection, RNA extraction and RT-PCR
• High sensitivity and high specificity – Limit of detection (LoD): 1 copy/µl (5 copies/reaction)
• Simple and easy-to-use procedure
• Accurate diagnosis regardless of SARS-CoV-2 variants (Alpha, Beta, Gamma, Delta, Omicron)
• The result report is sent to the laboratory by providing the reporting system (viewer program)

Product Principle

• real-time RT-PCR
• Target genes: RdRp gene and N gene
• Kit includes human targeted primer and probe set RNase P gene that serves as an internal positive control for the validation of complete procedures, including sample collection, RNA purification and PCR

Presentation Features

1. Analytical Sensitivity (Detection limit)

• Limit of detection (LoD) : 1 copy/µl (5 copies/reaction)

2. Precision

• Repeatability within CV 3%
• Reproducibility within CV 2%

3. Transfer Prevention pollution

• Prevention of transfer contamination by use of UDG

4. Analytical Specificity (cross-reactivity)

• No cross-reactivity with 26 tested respiratory pathogens

5. Analytical Specificity (interference)

• No interference with 9 tested interfering substances

SARS-CoV-2 virus variant detection test

• Kit used: PaxView® SARS-CoV-2 real-time RT-PCR kit
• Equipment Used: CFX96 (Bio-Rad)
• The specimen used: SARS-CoV-2, SARS-CoV-2 variants (alpha, beta, gamma, delta)
• Specimen source: NCCP (National Culture Collection of Pathogens) of Korea Disease Control and Prevention Agency
• Test method: test with a 1/10 serial dilution of each virus