Human ALK fusion gene detection primer set and detection kit

Abstract

The invention discloses a human ALK fusion gene detection primer set and a detection kit. The primer set comprises one or more detection primers in sixteen ALK fusion gene types A01 to A16 and Taqman probes corresponding to the gene types. As many as sixteen fusion gene types except for EML4 can be detected through the detection kit formed by the primer set and the probes, the requirement for the sample RNA obtaining quality is low, and the ideal detection effect can be achieved through samples in different existence forms; by means of the kit, an operator only needs to directly feed extracted sample RNA once, inverse transcription and PCR amplification are carried out in one closed tube reaction system, and the pollution probability and the result error probability are reduced. One-time detection only takes eighty minutes. The human ALK fusion gene detection primer set and the detection kit have the remarkable advantages that the number of the detection types is large, sensitivity is high, experiment operation is simple, the period is short, the human ALK fusion gene detection primer set and the detection kit are safe and nontoxic, and cost is low.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and clinical molecular diagnosis, in particular to primers, probes and kits for detection of human ALK fusion gene. Background technique [0002] Lung cancer is the most common malignant tumor in the world, and 80%-85% of the cases are non-small cell lung cancer (NSCLC). Both the morbidity and mortality of lung cancer in China are increasing significantly. According to the statistics of the Ministry of Health of my country, the death rate related to lung cancer ranks first in the total death rate of malignant tumors. Surgery, radiotherapy, and chemotherapy have been the mainstays of treatment for NSCLC. In recent years, with the deepening of research on tumor pathogenesis and its biological behavior, the field of molecular targeted therapy for lung cancer has become a research hotspot. ALK (anaplastic lymphoma kinase anaplastic lymphomakinase) rearrangement is found in a subtype of anapl...

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Problems solved by technology

[0005] The significant advantage of FISH is that it can specifically detect ALK recombination, and its probes have been commercialized, but there are many deficiencies in meeting the current clinical needs, such as: the specific fusion variant of the ALK fusion gene cannot be clarified; patients with advanced NSCLC usually can only provide about 2 mm For small biopsy tissues, it is difficult to ensure that there are more than 50 cancer cells in each field of view for result interpretation; for FFPE tissues, it is difficult to distinguish the tissue structure and shape under the microscope; the requirements for operation and interpretation techniques are very high, and doctors with rich operating experience can judge the results It is reliable, technology promotion is difficult and the cost of detection is expensive

In view of the above reasons, FISH detection is not suitable for large-scale screening and diagnosis of ALK-positive NSCLC patients at this stage

[0006] Monoclonal antibody combined with immunohistochemical method to detect the expression status of ALK has the advantage of being simple and fast, and has screening value for samples such as lung cancer histology and cytology, but it also needs to consider the sensitivity of immunohistochemical method and the determination of negative and positive always exists Certain subjectivity, the interpretation of weak positive results requires further confirmation by FISH or sequence-specific PCR techniques

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