Primer set for detecting oocyst deoxyribonucleic acid (DNA) of Cryptosporidium by using loop-mediated isothermal amplification technology, kit, detection method and application

A ring-mediated isothermal and technical detection technology, applied in the detection of Cryptosporidium oocyst DNA, the primer set and application of detection of Cryptosporidium oocyst DNA, the field of detection of Cryptosporidium oocyst DNA by warm amplification technology, can solve Complicated principles and other issues, to achieve the effect of wide application prospects, strong specificity, and simple operation

Inactive Publication Date: 2011-08-31
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Claims
  • Application Information

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Problems solved by technology

Although LAMP has high requirements for primer design, the principle is relatively complicated, and the actual

Method used

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  • Primer set for detecting oocyst deoxyribonucleic acid (DNA) of Cryptosporidium by using loop-mediated isothermal amplification technology, kit, detection method and application
  • Primer set for detecting oocyst deoxyribonucleic acid (DNA) of Cryptosporidium by using loop-mediated isothermal amplification technology, kit, detection method and application
  • Primer set for detecting oocyst deoxyribonucleic acid (DNA) of Cryptosporidium by using loop-mediated isothermal amplification technology, kit, detection method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0027] 1. Materials

[0028] 1.1 Insect strain (oocyst)

[0029] The improved acid-fast staining method was used to determine the natural infection of Cryptosporidium oocysts from cattle. The feces were collected from a cattle farm in Xuzhou for method establishment; other positive samples were collected from cattle and pigs in Shanghai, Xuzhou, Shaoxing, Harbin, Zhengzhou, and Aba, Sichuan. , sheep, dogs and other animals and people; on-site samples were collected from Shanghai, Shaoxing and Sichuan Aba cattle, pigs, sheep and other animals and people. In this example, 47 Cryptosporidium oocyst DNA-positive samples, 50 Cryptosporidium oocyst DNA-negative samples and 50 stool DNA samples collected from different sources were used. The human Giardia lamblia trophozoites used in this experiment were donated by Capital Medical University and stored in liquid nitrogen.

[0030] 1.2 Main reagents and instruments

[0031] Bst DNA polymerase was purchased from Biolabs in the UK; B...

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Abstract

The invention discloses a primer set for detecting oocyst deoxyribonucleic acid (DNA) of Cryptosporidium by using a loop-mediated isothermal amplification technology. The primer set comprises an external primer pair (F3,B3) and an internal primer pair (FIP,BIP), wherein F3 has a sequence shown in SEQ ID No: 1 or a complementary strand of the sequence; B3 has a sequence shown in SEQ ID No: 2 or a complementary strand of the sequence; FIP has a sequence shown in SEQ ID No: 3 or a complementary strand of the sequence; and BIP has a sequence shown in SEQ ID No: 4 or a complementary strand of the sequence. The invention also discloses a method for detecting the oocyst DNA of the Cryptosporidium by performing a loop-mediated isothermal amplification reaction by using the primer set, a kit for detecting the oocyst DNA of the Cryptosporidium, and application of the primer set to the preparation of a detection agent of the oocyst DNA of the Cryptosporidium. In the invention, the rapid detection of the oocyst DNA of the Cryptosporidium is realized by designing two pairs of specific primers of the Cryptosporidium, so that the speed and the accuracy rate of clinical diagnosis of cryptosporidiosis are improved.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a method for detecting Cryptosporidium oocyst DNA, in particular to a method for detecting Cryptosporidium oocyst DNA using a loop-mediated isothermal amplification technique; in addition, the present invention also relates to using Primer set and application of loop-mediated isothermal amplification technique for detection of Cryptosporidium oocyst DNA. Background technique [0002] Cryptosporidium (Cryptosporidium) is a serious opportunistic pathogenic zoonotic gastrointestinal protozoa. Oocysts pollute water sources and food, causing outbreaks of cryptosporidiosis. It has a wide range of hosts and can infect humans. More than 240 kinds of animals. Cryptosporidium is distributed globally, and its infection cases have spread to more than 90 countries on 6 continents. It causes diarrhea after infecting humans and animals, and is one of the main causes of death of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCY02A50/30
Inventor 沈玉娟袁忠英曹建平尹建海徐馀信
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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