Method for directly amplifying single ovum DNA (Deoxyribonucleic Acid) of human ascarid without extraction

Abstract

A method for directly amplifying the DNA of a single human roundworm egg without extraction, comprising four steps of washing the egg with distilled water, breaking the egg, PCR amplification and result detection. The invention can obtain PCR products with good stability and high amplification efficiency by optimizing the fragmentation method of eggs and the PCR amplification system, and can avoid the amplification failure caused by the loss of DNA in the extraction process. The amplified product obtained by the present invention can effectively perform downstream gene sequencing and STR scanning, and provide important technical methods for molecular epidemiology, genetic diversity, genetic structure, phylogenetics, phylogenetic geography and paleontological research of human roundworms , the invention is also of great significance to the molecular diagnosis, prevention and control of human ascariasis.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to a method for amplifying the DNA of a human ascaris single egg. Background technique [0002] human roundworm ( Ascaris lumbricoides Linnaeus, 1758), belonging to the phylum Nematodes, Class Caudosensilla, Order Ascariae, and the genus Ascaris. It is one of the most common intestinal parasites in the human body, especially children. It parasitizes in the human small intestine and causes ascariasis. It is a large human parasitic nematode. Mature females and males mate and lay eggs in the small intestine. The fertilized egg is excreted with the patient's feces and develops under suitable conditions. It can make people infected with ascariasis, which is called infectious egg. If this infectious egg enters the human body along with food or other ways, the larvae hatch in the duodenum, and the larvae drill into the blood vessels of the intestinal wall, pass through the liver, heart, lungs...

AI Technical Summary

Problems solved by technology

[0003] For the direct amplification of any kind of trace DNA, the release of genomic DNA is one of the key factors to determine whether the DNA amplification is successful. The special feature of human roundworm eggs is that the roundworm eggs have a layer of hard, dense protein The shell and the impermeable lipid layer make the eggs highly resistant to various physical and chemical substances and adverse external environments, but have a certain impact on the release of genomic DNA from the eggs

At the same time, the genome DNA content of human roundworm eggs is very small, so it is very difficult to extract DNA

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