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OFR1ab / N Dual Nucleic Acid Detection Kit (RT-qPCR)

Description:Novel pneumonia (NCP) is a respiratory infection caused by a new type of virus that was first identified in late 2019.This kit utilizes fluorescence quantitative probe-based PCR and guarantee a high specificity to ensure accurate one-step identification of ORF1ab and N genes. The kit offers a highly sensitive test with a limit of detection as low as 500 copies/ml. The results are available within 1.5 hour.Kit Contents:

Note: The kit components of different lots can not be interchanged.Storage and Transportation:Store at -20ºC±5ºC. Can be stored at 4 ° C for a short period of time. Shelf life 1 year.Repeated freezing and thawing ≤5 times.Protect from light.Applicable equipment:ABI series, Bio-Rad, Roche series and other fluorescent real-time PCR instruments.Specimens requirements:Specimen type: nasoryngeal swab from suspected infection; virus culture fluid, etc.2. Specimen collection:ryngeal, respiratory samples. Use a cotton swab to take secretions sample. Place the swabs in a centrifuge tube containing 3.0ml Virus preservation fluid and send them for inspection immediately. Basic Protocol:

Take the test sample and extract the RNA nucleic acid according to the instructions of the nucleic acid extraction kit. The nucleic acid extraction product should be stored at -20 ° C.

According to the total of test samples, the of PCR reaction tubes needed is "N". N = of samples + 1 negative control + 1 positive control.The following protocol is recommended for a 20 µl reaction volume. If the volume of reaction changes, please adjust proportionally.

Add 5 µl each of the extracted RNA, positive and of negative control to corresponding reaction tubes. After assembling all the components, cover the tube caps and gently mix the contents of the tube, mix well, and centrifuge briefly.

Place the reaction tube inside a real-time PCR instrument.Set the channel and sample information, reaction system volume 25 µl.Select the following channels: FAM channel for ORF1ab, VIC channel for N.Perform quantitative PCR using recommended cycling parameters settings:

Result analysis:1) Set the baseerally, it is set to 6-15cycle for ABI 7500, 7700 and other instruments, 3-15cycle for PE5700, and 6-12cycle for MJ Research Option2. Under special circumstances, the basebe adjusted accordantly.2) Set the threshold: The threshold exceeds the highest point of the negative control amplification curve (random noise pan>Result determination:1) Positive: The Ct value of the two channels of the test sample is ≤ 38.0, and the curve has a significant exponential growth period, the result is determined to be positive;2) Suspicious: Any test sample with a Ct value > 38.0 and < 40.0, repeat the experiment, if the Ct value is still < 40.0, and the curve has a significant exponential growth period, it is positive, otherwise it is negative;3) Negative: If the Ct value is not detected or the Ct value is >40, the result is negative.Quality control standards:1) Negative quality control: no logarithmic growth se or Ct value display in the amplification curve;2) Positive quality control: the amplification curve has a significant logarithmic growth se, and the Ct value is ≤30;The above conditions should be met at the same time, otherwise the experiment is deemed invalid.Limitations of detection method:

Precautions:

About AIVD BiotechEstablished in 2014, AIVD Biotech Inc. is a biotech company, located in Shenzhen, China, a ISO 13485, ISO 9001 certificated enterprise.AIVD is a leading manufacturer of solutions for immunodiagnostics.Our company develops and produces protein raw materials: such as antibodies, antigens for immunoassay development; uncut sheets, POCT rapid tests kits, molecular diagnostics and offers various custom services for In Vitro diagnostic industry.