Application of protoporphyrin in preparation of diagnostic kit for hemocytogenic protoporphyrinosis

Abstract

The invention provides an application of protoporphyrin in preparation of a diagnostic kit for hemocytogenic protoporphyrinosis. According to the diagnostic kit, the excitation spectrum of a fluorescence detection whole blood sample, the emission spectrum of protoporphyrin and zinc porphyrin and the fluorescence intensity of protoporphyrin and zinc porphyrin are used as diagnostic parameters respectively, and according to the wave crest conditions of the excitation spectrum and the emission spectrum, the detection sensitivity of the whole blood sample is improved. And whether the protoporphyrin is abnormally accumulated according to the comparison result of the fluorescence intensity of the protoporphyrin and the fluorescence intensity of the zinc porphyrin is taken as a diagnosis threshold value, so that whether the ferrous chelating enzyme coding gene is mutated is judged, and the diagnosis kit greatly improves the mutation detection rate of the ferrous chelating enzyme coding gene, and is simple, convenient, direct and rapid.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of protoporphyrin in the preparation of a diagnostic kit for hematopoietic protoporphyria. Background technique [0002] Erythropoietic protoporphyria (EPP) is a genetic disease that results in heme synthesis disorders due to mutations in the gene encoding ferrochelatase. Therefore, the presence of EPP is usually determined by detecting the mutation of the gene encoding ferrochelatase, but the carrier rate of the gene mutation of ferrochelatase in EPP patients is extremely low, and the actual detection rate is even lower. Since mutations in the ferrochelatase gene can lead to abnormal accumulation of erythrocyte protoporphyrin (especially protoporphyrin not bound to metal atoms) in the blood, the presence of protoporphyrin can also be judged indirectly by detecting the abnormal accumulation of protoporphyrin. For ferrochelatase gene mutation, commonly used detection ...

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Problems solved by technology

However, the above methods either have low accuracy or are too complicated to be widely used due to the high cost.

Fluorescence detection method is a type of detection method that has been studied more. However, although protoporphyrin has the characteristic of being excited by a specific wavelength, body fluid specimens not only contain a single substance, protoporphyrin, but usually also contain many kinds of organic substances. / inorganic substances, such as bilirubin, hemoglobin, etc., and these organic / inorganic substances each have unique fluorescence characteristics, therefore, direct use of fluorescence spectroscopy to detect protoporphyrin in body fluid samples, in other inorganic / organic substances Spectra may partially or completely overlap under the influence of

It is also recognized in the world that there are many limitations in the various detection methods of protoporphyrin, which greatly affect the popularization of applications (Gou, E.W., et al., Pitfalls in Erythrocyte ProtoporphyrinMeasurement for Diagnosis and Monitoring of Protoporphyrias. Clin Chem, 2015.61 (12) :p.1453-6.)

If the detection of protoporphyrin is abandoned and the gene detection is carried out directly, according to the reported extremely low prevalence rate (ranging from 1:75,000 to 1:200,000) (Elder, G., et al., The incidence of inherited porphyrias in Europe.J Inherit Metab Dis,2013.36(5):p.849-57.), extremely low cost performance

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