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Cat.No: BT-QCY-0093 Datasheet Instruction for Use
Specification Quantities
Price $1839.72–3066.20
| Product Name | Mycobacterium avium PCR Kit |
| Catalog No. | BT-QCY-0093 |
| Description | Mycobacterium avium belongs to the genus Mycobacterium within the family Mycobacteriaceae and is a Gram-positive bacterium. Mycobacterium avium is an environmental saprophytic bacterium that primarily colonizes birds and their bodies, and can be transmitted via the respiratory tract, digestive tract, and mucous membranes. Following infection, Mycobacterium avium can affect various tissues and organs, including the lungs, bone marrow, and lymph nodes. Clinical manifestations primarily include four types: solitary nodules, nodular bronchiectasis, nodular infiltrates, and diffuse infiltrates in immunocompromised patients. Mycobacterium avium is a zoonotic disease; if left untreated, it can lead to respiratory failure and may even be life-threatening. Therefore, rapid detection of Mycobacterium avium is of great significance. |
| Features | Ready-to-use; users only need to provide a DNA template sample. The primers and probes have been optimized to ensure high analytical sensitivity, capable of detecting as few as 100 copies per reaction. A positive control is provided to help distinguish false-negative samples. High specificity: The primers are designed based on highly conserved regions of Mycoplasma DNA and will not cross-react with DNA from other organisms. Suitable for both qualitative and quantitative detection. The linear range for quantitative detection is at least five orders of magnitude. This product is sufficient for 50 probe-based qPCR reactions in a 20 μL system. |
| Kit Components | 2× ProbeqPCR Master Mix: 0.5 mL Fluorescent PCR-specific template diluent: 1 mL Ultrapure water: 1 mL Mycobacterium avium qPCR primer-probe powder: 50 reactions Mycobacterium avium qPCR positive control (1E7 copies/μL): 50 μL User manual: 1 copy |
| Assay Method | PCR |
| Sample Preparation | If there are N samples, it is recommended to prepare N+2 extractions: one extra for the PC (positive control for sample preparation) and one for the NC (negative control for sample preparation). Use 10 μL of the fourth dilution solution obtained in the previous step, add an appropriate amount of water to achieve a total volume matching the required initial sample volume for each sample preparation, and use this as the PC. Use water as the NC. Purify the sample DNA using a method of your choice. |
| Test Procedure | Since the standard samples are highly concentrated, the following dilution procedures must be performed in a separate area to avoid contaminating the samples or other components of this kit. To ensure product stability and prevent the spread of infectious pathogens, this product does not include live samples as positive controls; instead, it provides non-infectious DNA fragments as positive controls. Label 6 centrifuge tubes with the numbers 6, 5, 4, 3, 2, and 1. Using a pipette tip with a filter (use filter-tipped pipette tips; the same applies below), add 45 μL of the dedicated fluorescent PCR template dilution buffer to each tube. Add 5 μL of the 1E7 copies/μL positive control (provided in the kit) to tube 6. Vortex thoroughly for 1 minute to obtain a 1E6 copies/μL standard curve sample. Place on ice until ready for use. Change the pipette tip. Add 5 μL of the 1E6 copies/μL positive control (prepared in the previous step) to tube 5. Vortex thoroughly for 1 minute to obtain a 1E5 copies/μL standard curve sample. Place on ice until needed. Change the pipette tip. Add 5 μL of the 1E5 copies/μL positive control (prepared in the previous step) to tube 4. Vortex thoroughly for 1 minute to obtain a standard curve sample at 1E4 copies/μL. Place on ice until needed. Repeat the above procedure until 6 standard curve samples at different dilutions are obtained. Place on ice until needed. If performing quantitative analysis with only one replicate, label N+9 PCR tubes: N+2 for the N+2 samples obtained in the previous step, 1 for the PCR negative control (using water as the template), and 6 for the standard curve. If performing qualitative analysis with a single replicate, label N+4 PCR tubes: N+2 for the N+2 samples obtained in the previous step, 1 for the PCR negative control (using water as template), and 1 for the PCR positive control. |
| Results | If the positive control from sample preparation or the PCR positive control (including the standard curve sample) yields a negative result, the entire experiment is invalid. There is no need to analyze the data; sample preparation must be repeated, and PCR amplification must be performed again. If the negative control from sample preparation or the PCR negative control yields a positive result, indicating environmental contamination, the entire experiment is invalid. There is no need to analyze the data; contact the manufacturer. If both the negative control and the positive control are normal, the experiment is valid and may proceed to subsequent analysis. If this kit is used for quantitative detection, plot a standard curve with the log value of the positive control concentration on the x-axis and the Ct value on the y-axis. Then, use the Ct value of the sample to calculate the log value of the sample’s DNA concentration from the standard curve, and subsequently determine its concentration. If this kit is used for qualitative testing (to determine only positive or negative results), the negative control must have no Ct reading or a Ct value greater than or equal to 40. The positive control must exhibit a logarithmic increase in fluorescence, a typical amplification curve, and a Ct value less than 40. For test samples, if the Ct value has no reading or is greater than or equal to 40, the result is negative; if it is less than 40, the result is positive. |
| Storage and Transportation | Transport at low temperatures; store at -20°C. Shelf life: 2 years |
For research use only, not for clinical use.
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