Primer, detection kit and detection method for screening SCID genetic diseases

Abstract

The invention relates to a primer, detection kit and detection method for screening SCID genetic diseases. The kit comprises a DNA extract, a PCR MIX reaction solution and upstream and downstream primers and probes for TREC gene amplification, internal reference Calpha gene amplification and external control Calpha gene amplification; the sequences of the upstream and downstream primers for TREC gene amplification are shown as SEQ ID NO.1 and SEQ ID NO.2, and the sequence of the TREC gene probe is shown as SEQ ID NO.3; the sequences of the upstream and downstream primers for the internal reference Calpha gene amplification are shown as SEQ ID NO.4 and SEQ ID NO.5, and the sequence of the internal reference Calpha gene probe is shown as SEQ ID NO.6; the sequences of the upstream and downstream primers for the external control Calpha gene amplification are shown as SEQ ID NO.4 and SEQ ID NO.5, and the sequence of the external control Calpha gene probe is shown as SEQ ID NO.7. The methodis based on a fluorogenic quantitative PCR technology. A chelex100 method is used for dealing with dried blood spots for direct amplification to qualitatively detect T cell receptor deletion loops, and a rapid, efficient and sensitive SCID screening technology system is established. The detection method is convenient in operation, low in cost and easy to popularize.

Description

technical field [0001] The invention belongs to the field of gene screening for genetic diseases of newborns, and in particular relates to a primer, a detection kit and a detection method for screening of SCID genetic diseases. Background technique [0002] Severe combined immunodeficiency disease (SCID) is a deficiency and / or dysfunction of T lymphocytes, B lymphocytes, and NK cells caused by multiple genetic factors, resulting in serious defects in both humoral immunity and cellular immunity. This primary immunodeficiency is often defined as a lack of naive T cells. The incidence of SCID is about 1 / 100,000 live births, but because children with SCID often die before a definite diagnosis, this number may be seriously underestimated. Without treatment, the case fatality rate of SCID is 100%. 95% of children are boys, polymorphic linked recessive inheritance, autosomal recessive inheritance and sporadic cases occasionally. The clinical manifestations are that children have ...

AI Technical Summary

Problems solved by technology

However, since SCID patients lack clinical symptoms before infection, how to identify early before infection becomes the key

[0005] At present, the United States and other developed countries and my country's Taiwan region have carried out newborn SCID screening work, and the US Centers for Disease Control and Prevention has also included this project in the external quality evaluation system for newborn disease screening, but domestic SCID newborn screening And the corresponding genetic diagnosis and testing work is still in the preliminary stage

At present, there is no report on large-scale SCID screening of neonatal population in China, and no clear neonatal SCID screening technology system has been established. Therefore, it is of great significance to establish a fast, efficient and sensitive SCID screening technology system

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