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Human EML4-ALK Fusion Gene Mutation Detection Kit (Fluorescence PCR)

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Human EML4-ALK Fusion Gene Mutation Detection Kit (Fluorescence PCR)

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Introduction

[Product Name] Human EML4-ALK Fusion Gene Mutation Detection Kit(Fluorescence PCR)

[Packaging Size] 20 tests/kit, 50 tests/kit
[Intended Use]
This kit is used to qualitatively detect 12 mutation types of EML4-ALK fusion gene in samples of human nonsmall cell lung cancer patients in vitro (Table 1). The test results are for clinical reference only and should not be used as the sole basis for individualized treatment of patients. Clinicians should make comprehensive judgments on the test results based on factors such as the patient's condition, drug indications, treatment response, and other laboratory test indicators.
Lung cancer is the most common malignant tumor worldwide, and 80%~85% of the cases are nonsmall cell lung cancer (NSCLC). Gene fusion of echinoderm microtubule-associated protein-like 4 (EML4) and anaplastic lymphoma kinase (ALK) is a novel target in NSCLC, EML4 and ALK are respectively located in human the P21 and P23 bands on chromosome 2 and are separated by approximately 12.7 million base pairs [1]. At least 20 fusion variants have been found, among which the 12 fusion mutants in Table 1 are common, where mutant 1 (E13; A20) is the most common one, followed by mutants 3a and 3b (E6; A20), accounting for about 33% and 29% of patients with EML4-ALK fusion gene NSCLC, respectively [2]. ALK inhibitors represented by Crizotinib are small-molecule targeted drugs developed for ALK gene fusion mutations. By inhibiting the activity of the ALK tyrosine kinase region, blocking its downstream abnormal signaling pathways, thereby inhibiting the growth of tumor cells , to achieve targeted therapy for tumors [3]. Clinical studies have shown that Crizotinib has an effective rate of more than 61% in patients with EML4-ALK fusion mutations, while it has almost no effect on wild-type patients [4]. Therefore, the detection of EML4-ALK fusion mutation is the premise and basis for guiding the use of Crizotinib drugs. This test kit is used to guide the administration of ALK inhibitors and provide the basis for personalized medicine for patients with nonsmall cell lung cancer.

[Test Principles]

This kit uses fluorescent probe real-time PCR technology to design specific primer probes for fluorescent detection for the target sequence of 12 fusion mutations of the EML4-ALK fusion gene and the internal control gene. The 5' of the specific probe is labeled with a FAM fluorophore (12 fusion mutations) and VIC (HEX) fluorophore (internal control), 3' is labeled with a BHQ1 quencher. The extracted RNA is reverse transcribed into cDNA. During the PCR amplification process, specific primers and probes bind to their respective target sequences. When Taq enzyme encounters the probes bound to the target sequence, it exerts the function of 5′-end exonuclease, so that the reporter and the quencher is separated, and the fluorescence monitoring system can receive the fluorescent signal, that is, each time a DNA chain is amplified, a fluorescent molecule is formed, realizing the complete synchronization of the accumulation of the fluorescent signal and the formation of the PCR product. The qualitative detection of 12 fusion mutations in the sample and the detection of the internal control are realized by two reaction buffer.

At the same time, the endogenous internal control is introduced as the quality control of reagents, RNA quality and the operation itself. Even if the RNA is degraded, the internal control can truly reflect the effective RNA amount in the system.

[Storage Conditions and Shelf-life]

The kit should be stored below -18°C protected from light. The shelf life is 9 months(please use within the validity period). After opening, the number of repeated freezing and thawing should not be more than 4 cycles. It can be stably stored for 5 days below -18°C and protected from light during transportation.

See the packaging label for the production date 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.

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