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HWTS-GE013A Human Leukocyte Antigen B27 Nucleic Acid Detection Kit (Fluorescence PCR )

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HWTS-GE013A Human Leukocyte Antigen B27 Nucleic Acid Detection Kit (Fluorescence PCR )

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Introduction

[REF] HWTS-GE013A

[Specification] 50 tests/kit

[Research Use Only]

This kit is suitable for qualitative detection of the DNA in the human leukocyte antigen subtypes HLA-B*2702, HLA-B*2704 and HLA-B*2705.

This kit can be used as an aid for ankylosing spondylitis detection. Ankylosing spondylitis (AS) is a chronic progressive inflammatory disease that mainly affects the spine and can involve the sacroiliac joints and surrounding joints to varying degrees. It has been revealed that AS exhibits obvious familial aggregation and is closely related to human leukocyte antigen HLA-B27. In humans, more than 70 HLA-B27 subtypes have been discovered and identified, and of them, HLA-B*2702, HLA-B*2704 and HLA-B*2705 are the most common subtypes related to the disease [1-10]. In China, Singapore, Japan and Taiwan district, HLA-B27 is most common and accounts for approximately 54%, followed by HLA-B*2705 which accounts for 41% [1-4]. This kit can detect the DNA in HLA-B*2702, HLA-B*2704 and HLA-B*2705, but does not differentiate them from each other.

Experimental operators should have received professional training in gene amplification or molecular biologic testing, and have qualifications relevant to experimental operation. The laboratory should be equipped with rational biological safety precautions and protective procedures.

[Test Principle]

This kit uses fluorescence-based PCR for qualitative detection of the DNA in the subtypes HLA-B*2702, HLA-B*2704 and HLA-B*2705. The HLA-B27 gene is located on the short arm of human chromosome 6. So far, more than 70 HLA-B27 subtypes have been discovered and identified, and there are differences in gene sequence among these subtypes. This kit contains primers and probes specific for HLA-B*2702, HLA-B*2704 and HLA-B*2705 in exon 3; its internal control is designed in the conserved region of the Globin gene and serves as a non-competitive internal standard that can monitor the entire experimental process. This kit contains an enzyme system composed of dUTP/UNG, which can effectively prevent environmental contamination. In the kit, the target gene detection probe is labeled with the fluorophore FAM, and the internal control gene detection probe is labeled with the fluorophore VIC.

[Storage Conditions and Shelf Life]

This kit should be stored at -18°C protected from light. Its shelf life is 18 months. It should be transported using airtight  foam boxes with dry ice or ice bags. After opening, the kit should be temporarily stored at 4°C protected from light for no more than 15 days (validation has not been performed for storage for > 15 days); it is recommended not to store the kit at 37°C for more than 5 days; the kit shall not be subjected to repeated freezing and thawing for more than 5 cycles (validation has not been performed for > 5 cycles), and given that the enzyme should not be subjected to repeated freezing and thawing, it is recommended that after opening, the kit be used up once for all, or the remaining product be stored at 4°C. Repeated freezing and thawing is prohibited.

[Applicable Equipment]

ABI 7500, ABI Stepone, Roche LightCycler 480, Stratagene MX 3000P Real-time Fluorescence-based qPCR Instrument, Ultrassay XP96 Real Time qPCR System.

[Acceptable Specimens]

1. Samples are collected from confirmed or suspected AS patients. Samples should be collected aseptically.

2. Collected samples are whole blood samples from confirmed or suspected AS patients. The sample volume is 0.5 mL, and the anticoagulant EDTA is added to each sample. A label with a unique identification number is affixed to the exterior of each tube. Repeated freezing and thawing of samples should be avoided during transportation and storage. If the condition -20°C cannot be ensured, samples should be transported at 0-8°C. The samples may be stored at -20°C for 6-8 months and below -70°C for a longer time. Necessary information such as the sample number, the date of disease onset, and the date of sample collection, should be attached during transportation and storage.

3. For whole blood samples stored at 4°C for 5-6 months, the detection accuracy can reach more than 95%.

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