COVID-19 FluA&B real time PCR detection (100 tests) - Research Use Only

COVID-19 FluA&B real time PCR detection (100 tests)

Cat. No.: BMG-QP2178-01
Size: 100 tests


Description

The Influenza SARS-CoV-2 (Flu SC2) Multiplex Assay is a real-time RTPCR multiplexed test intended for the simultaneous qualitative detection and differentiation of SARS-CoV-2, influenza A virus, and/or influenza B virus nucleic acid in upper or lower respiratory specimens (such  as nasopharyngeal, oropharyngeal and nasal swabs, sputum, lower respiratory tract aspirates, bronchoalveolar lavage, and nasopharyngeal  wash/aspirate or nasal aspirate) collected from individuals suspected of respiratory viral infection consistent with COVID-19 by a healthcare  provider1. Symptoms of respiratory viral infection due to SARS-CoV-2 and influenza can be similar. Test results must be combined with clinical  observations, patient history, and epidemiological information.

Summary and explanation of the test

The novel coronaviruses belong to the β genus. COVID-19 is an acute respiratory infectious disease. The Flu SC2 Multiplex Assay is a molecular in vitro diagnostic test that aids in the detection and differentiation of RNA from SARS-CoV-2, influenza A virus, and/or influenza B virus and is based on widely used nucleic acid amplification technology. The product contains oligonucleotide primers and dual-labeled hydrolysis probes (TaqMan®), and control material used in qRT-PCR for the in vitro qualitative detection and differentiation of SARS-CoV-2 virus RNA, influenza A virus RNA and/or influenza B virus RNA in upper and lower respiratory specimens.

Principle

The Flu SC2 Multiplex Assay is a real-time reverse transcription polymerase chain reaction (rRT-PCR) test. The Flu SC2 Multiplex Assay contains three primer/probe sets (InfA, InfB, and SC2) that target the RNA of influenza A virus, influenza B virus, and SARS-CoV-2 virus, respectively. The assay also contains primers and a probe to detect the human RNase P gene (RP) in clinical specimens or control samples. The COVID-19 Real Time PCR Detection Kit also contains a positive control (Plasmid DNA) and a negative control for testing process monitoring.

Reagents and materials provided
 

Item	Component	Quantity		Volume /T
		100 Tests / Box	500 Tests / Box
1	PCR Mix	1.5 mL/ tube × 1	1.5 mL/ tube × 5	14 µL
2	RT-PCR Enzyme Mix	110 μL/ tube × 1	650 μL/ tube × 1	1 µL
3	Positive Control	50 μL/ tube × 1	250 μL/ tube × 1	5 μL
4	Negative Control	50 μL/ tube × 1	250 μL/ tube × 1	5 μL

Storage instructions

Store the kit frozen at -20±5°C and avoid repeated freeze-thaw cycles (less than 5 frozen-thaw times allowed). The product is valid for 12 months, and the expiry date of the product is shown on the label.

Specimen type and preservation

Specimen type: oropharyngeal swabs, nasopharyngeal swabs and sputum.
Specimen preservation: process specimens for viral RNA extraction and nucleic acid detection as soon as possible. Specimens can be stored at 4°C for 24 hours, or frozen at -80°C for longer storage, if not processed immediately. Avoid repeated freeze-thaw cycles during transport and storage of the specimens

Assay PCR reaction setup (at reagent preparation area)

A. Reagent Preparation
To calculate the number (n) of PCR reaction tubes (n= number of specimens to be tested + number of positive controls + number of negative controls), add n x14 μL of SARS-CoV-2 PCR Mix and n x 1 μL of RT-PCR Enzyme Mix to a clean tube, vortex to mix well, centrifuge and aliquot 15 µL of the mix into each PCR reaction tube.

B. Addition of Viral RNA
Add 5 μL of viral RNA extracts of the specimen or positive/negative control to each PCR tube according to the reaction preparation table below for testing. Then immediately cap the tube tightly, briefly spin the mixture and move it to the amplification area for RT-PCR detection

• PCR Reaction Preparation

C. RT-PCR amplification (at Amplification Area, use Applied Biosystems 7500 Real-Time PCR System as an example)

1. 1. Place the complete PCR reaction tubes into the fluorescent quantitative PCR analyzer and label positive control, negative control and testing specimen on the Thermocycler software.
2. 2. Enter the PCR reaction volume: 20 μL.
3. 3. Select fluorescence detection channels: JOE, FAM and Cy5 channels. Cy5 is the E gene indicator channel, FAM is the ORF1ab indicator channel, and JOE is the RNase P gene indicator channel (internal control).
4. 4. Select PCR cycle parameter setting following the instructions below, save the file and start the PCR run.
5. 5. Analyze the results according to your thermocycler manufacturer’s instructions. If the positive and negative control samples do not meet their respective requirements, the PCR run must be repeated.

Determination of results

1. The results can only be determined when the results of both controls meet their requirements.
2. Criterion for a positive specimen: a specimen can be reported as positive when FAM, Cy5 and JOE channels are all positive.
3. Criterion for a negative specimen: a specimen can be reported as negative when the JOE channel is positive and the ROX, FAM and Cy5 channels are negative.

Compatibility of real-time PCR thermocyclers
Product performance

Product analysis performance evaluation results:

1. 1. The analytical sensitivity of this kit is 200 copies/mL.
2. 2. Cross-reaction: no cross-reaction with other pathogens such as seasonal influenza A (H1N1) virus, novelinfluenza A (H1N1-2009) virus, influenza AH3N2, H5N1, H7N9,influenza B Yamagata,influenza B Victoria, RSV A, RSV B, parainfluenza I, parainfluenza II, parainfluenza III, adenovirus types 1, 2, 3, 4, 5, 7 & 55, enterovirus types A, B, C and D, hMPV (human metapneumovirus), EB virus, measles virus, human cytomegalovirus, rotavirus, norovirus, mumps virus, varicella zoster virus, mycoplasma pneumoniae, chlamydia pneumoniae, legionella, bordetella pertussis, haemophilus influenzae, staphylococcus aureus, streptococcus pneumoniae, streptococcus pyogenes, klebsiella pneumoniae, mycobacterium tuberculosis, aspergillus fumigatus, candida albicans, candida glabrata, cryptococcus neoformans, coronavirus (HKU1, OC43, NL63, 229E), SARS-CoV-1, MERS coronavirus.

REFERENCE

1. National Health Commission & State Administration of Traditional Chinese Medicine. Diagnosis and Treatment Protocol for Novel Coronavirus Pneumonia (Trial Version 7). [R]. March 3, 2020