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Mycobacterium Tuberculosis DNA Detection Kit(Fluorescence PCR)

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Category:Respiratory tract infection

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Mycobacterium Tuberculosis DNA Detection Kit(Fluorescence PCR)

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Introduction

Mycobacterium Tuberculosis DNA Detection Kit(Fluorescence PCR)
[Packaging Size] 20 tests/kit, 50 tests/kit
[Intended Use]
It is suitable for qualitative detection of Mycobacterium tuberculosis DNA in human clinical sputum samples, and is suitable for auxiliary diagnosis of Mycobacterium tuberculosis infection. The test results are for clinical reference only, and the final diagnosis should be comprehensively considered in close combination with other clinical indicators.

Mycobacterium culosis, referred to as Tubercle bacillus(TB), is a class of obligate aerobic bacteria with positive acid-fast staining. It is amastigote and has fimbriae and microcapsules but not forms spores. Its bacterial wall has neither teichoic acid of gram-positive bacteria, nor lipopolysaccharide of gram-negative bacteria. Mycobacterium tuberculosis, which is pathogenic to humans, is generally considered to be human, bovine, and African. Its pathogenicity may be related to the inflammation caused by the proliferation of bacteria in tissue cells, the toxicity of bacterial components and metabolites, and the immune damage to the bacterial components. Pathogenic substances are associated with capsules, lipids and proteins. Mycobacterium tuberculosis can invade susceptible organisms through the respiratory tract, digestive tract or skin injury, causing tuberculosis of various tissues and organs, of which the most common is pulmonary tuberculosis through the respiratory tract. It usually occurs in children, and presents with symptoms such as low-grade fever, night sweats, and a small amount of hemoptysis. Secondary infection is mainly manifested as low-grade fever, night sweats, and hemoptysis. Mostly it is long-term chronic disease, and a few are acute. Tuberculosis is one of the top ten causes of human death worldwide. In 2018, about 10 million people around the world were infected with Mycobacterium tuberculosis, of which about 1.6 million died.

IS6110 is an inserted nucleotide sequence discovered by Thierry et al. in 1990 in Mycobacterium tuberculosis. IS6110 is present in multiple copies of a sequence in the Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis, and BCG genes. IS6110 is relatively stable in the strains in which it exists, and will not change the IS6110 DNA fingerprints when the strains develop drug-resistant mutations. The 38KD protein antigen is a phosphate transporter and contains 6 epitopes targeted by monoclonal antibodies specific for Mycobacterium tuberculosis. The diagnostic specificity of the reported 38KD is 88%-100%.

[Test Principles]

This kit uses the methods of combination of PCR amplification and fluorescent probe, selects the specific conserved region of Mycobacterium tuberculosis IS6110 and 38KD antigen sequence as the detection target gene region, and designs specific primer and probes for fluorescent detection. The specific probes 5' is labeled FAM (IS6110 and 38KD), VIC (HEX) (internal control gene) fluorophore, and the 3' quencher is BHQ1. In the process of PCR amplification, specific primers and probes bind to their respective target sequences. During the amplification process, the formation of PCR products is achieved through the DNA polymerase activity and 5'-3' exonuclease activity of Taq enzyme. Meanwhile the accumulation of fluorescent signals is completely synchronized. And the qualitative detection of sample nucleic acid is realized through one reaction buffer.

[Storage Conditions and Shelf-life]

Storage condition: The kit should be stored below -18°C protected from light. The shelf life is 12 months. The number of repeated freezing and thawing should not be more than 4 cycles. After opening, it should be stored below -18°C. If it is stored 2~8°C, please use it within 1 week. Transportation: The kit is stable for 5 days in a shipping box containing dry ice.

See the packaging label for the production date, production batch number and expiration date.

[Applicable Instruments]

Applied Biosystems 7500 Real-Time PCR Systems, Applied Biosystems 7500 Fast Real-Time PCR Systems, QuantStudio®5 Real-Time PCR Systems, SLAN-96P Real-Time PCR Systems(Shanghai Hongshi Medical Technology Co., Ltd. ), LightCycler®480 Real-Time PCR system, LineGene 9600 Plus Real-Time PCR Detection System(FQD-96A, Bioer technology), MA-6000 Real-Time Quantitative Thermal Cycler (Suzhou Molarray Co., Ltd.) , BioRad CFX96 Real-Time PCR System, BioRad CFX Opus 96 Real-Time PCR System.

[Requirements for Samples]
1. Sample Collection
For those with less sputum, let the patient rinse his mouth after waking up in the morning, then take a few deep breaths, cough up the sputum from the lungs, and collect it in a clean container for testing as soon as possible. For those with much sputum, first ask the patient to cough lightly and cough out the old sputum retained in the trachea, and then take a deep breath several times to cough up fresh sputum from the depths of the lungs, and send it for testing immediately. If there is no sputum, you can go to the hospital for atomization inhalation to induce sputum, and then send it for testing.
2. Precautions for sample collection: Avoid contamination during sample collection, storage and transfer.
Sample safety: All samples are considered to be potentially infectious, and operations are performed in accordance with relevant national standards.
3.Storage
The sputum samples to be tested should not be stored at 2−8°C for more than 24 hours; stored at temperatures below −18°C for no more than three months; can store at temperatures below −70°C for long time. Repeated freezing and thawing should be avoided.

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