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Detection kit used for detecting echinococcus shiquicus (larvae)

A technology of Echinococcus shiqu and a detection kit is applied in the field of detection kits for Echinococcus shiqu, which can solve the problems of low sensitivity and specificity, poor specificity, complicated operation, etc. Fast and accurate etiological identification, simple operation effect

Active Publication Date: 2013-11-27
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has the disadvantages of time-consuming and laborious, low sensitivity and specificity, and also has the risk of infecting people and the surrounding environment
However, there are few molecular biological detection methods for this species, and there are problems of poor specificity, low sensitivity and cumbersome operation.

Method used

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  • Detection kit used for detecting echinococcus shiquicus (larvae)
  • Detection kit used for detecting echinococcus shiquicus (larvae)
  • Detection kit used for detecting echinococcus shiquicus (larvae)

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0020] 1) Preparation of positive DNA from Echinococcus shiqu

[0021] According to the instructions of the DNA extraction kit, the genomic DNA was extracted from the Echinococcus shiquense collected from the lung tissue of rabbits in the wild plateau, and the concentration of the prepared genomic DNA was measured, diluted to 10ng / mL with diluent, and stored at -20°C for later use .

[0022] 2) Preparation of reaction primer mixture

[0023]The primer mixture is prepared according to the ratio of FIP:BIP:F3:B3 of 8:8:1:1, see Table 1,

[0024]

[0025] 3) Refer to Table 2 for the LAMP reaction system,

[0026]

[0027] Mix the above mixture and centrifuge.

[0028] The reaction conditions were 62°C, 50 min at 80°C, 5 min at 4°C, and stored permanently. After the reaction, the LAMP product is mixed with the color developer at a volume ratio of 25:1, and it turns yellow-green. For the results, see figure 1 Middle left picture, while the negative one is yellowish, see...

specific example

[0030] The samples in the reaction system were added to the 9 reaction tubes according to the above ratio, and the DNA samples were Echinococcus granulosus G1 strain DNA, Echinococcus multilocularis NDA, Echinococcus shiquero DNA, Taenia alveolar DNA, soybean Taenia zoster DNA, Taenia polycephala DNA, Taenia zosterus DNA, Dipylidium caninum DNA and sterile water blank control. Mix gently and centrifuge briefly. Amplify in a water bath at 62°C for 50 minutes, and immediately transfer to an 80°C water bath for inactivation for 5 minutes. Take 5 μL of the amplification product, use TAE as the buffer, and detect it in 100-volt electrophoresis in 1.5% agarose gel (containing 0.5 μg / mL ethidium bromide). See the test results image 3 . Nucleic acid amplification was only completed in the reaction system using Echinococcus shiquense DNA as a template, indicating that the specificity of the detection primers was good.

[0031] The samples were added in the above proportions to 11 ...

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Abstract

The invention discloses a detection kit used for detecting an echinococcus shiquicus (larvae). The detection kit comprises four primers, namely SEQ ID No.1, SEQ ID No.2, SEQ ID No.3 and SEQ ID No.4, or comprises a primer reaction mixture formed by the four primers. An LAMP method established according to the invention can overcome the defects of time and energy waste, low sensitivity and low specificity in the prior art, can realize detection without special instruments, equipment and long-time direct contact and only with a general water bath kettle, has the advantages of high sensitivity, good specificity and high sensibility, and is sensitive and fast in detection reaction and simple in operation, thereby being suitable for fast and accurate etiological identification in clinical application and laboratories.

Description

technical field [0001] The invention relates to a detection kit for detecting whether there are parasites in a sample to be inspected, in particular to a detection kit for detecting echinococcus shiqu (larvae). Background technique [0002] Echinococcosis, also known as hydatid disease, is caused by the genus Echinococcus ( Echinococcus ) Tapeworm larvae—echinococcus or hydatid cysts parasitize in animals (including humans), causing a type of zoonotic parasitic disease, which not only seriously endangers human health, but also hinders the development of animal husbandry, causing annual The economic loss is hundreds of millions of yuan. The disease is distributed worldwide, and my country is the main high-incidence area. Echinococcus stone canal ( Echinococcus .s hiquicus , Es) is a newly discovered species of Echinococcus, named after it was first discovered in Shiqu, Sichuan in 2005. This tapeworm is similar in shape to Echinococcus multilocularis in the genus Echinoc...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 贾万忠倪兴维娄忠子闫鸿斌李宏民李立
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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