Kit for detecting equine piroplasmosis and preparation method and using method thereof

A kit and the technology of Piroplasma, which are applied in the field of detection kits for the differential diagnosis of whether equine animals are infected with Piroplasma, can solve the problems of low detection rate and very high quality requirements for testing personnel, and achieve sensitivity and specificity High performance and extended detection range

Active Publication Date: 2011-09-14
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

At present, the detection and diagnosis of equine piriplasmosis mainly include the following types of means: (1) Traditional methods such as blood smear staining microscope detection

Method used

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  • Kit for detecting equine piroplasmosis and preparation method and using method thereof
  • Kit for detecting equine piroplasmosis and preparation method and using method thereof
  • Kit for detecting equine piroplasmosis and preparation method and using method thereof

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preparation example Construction

[0047] One, be used to detect the preparation process of the detection membrane of horse Piroplasma:

[0048] Target gene amplified based on Piroplasma universal primers (RLB-F / RLB-R)——Piroplasma 18S rRNA gene V 4 For hypervariable region sequences, the general DNA probes, genus-specific DNA probes (2 types), and species-specific DNA probes (2 types) of Piroplasma were designed using professional software. After the probe is artificially synthesized and modified with amino groups, the nucleic acid probe can be covalently bound to the negatively charged nylon membrane ( 0.45μm) on the membrane.

[0049] The specific operation procedure is as follows:

[0050] 1. Synthesis and modification of specific DNA probes for detection of Piroplasma

[0051] The general DNA probe C-all of Piroplasma, the general DNA probe B-all of Babesia, the general DNA probe T-all of Theileria, and the specific DNA probe of Babesia were synthesized respectively as described in Table 1. B-cab and t...

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Abstract

The invention discloses a detection kit for identifying and diagnosing whether an equine animal is infected with piroplasmosis or not, and a preparation method and a using method thereof. A detection film for detecting equine piroplasmosis and a piroplasmosis universal primer are arranged in the kit for detecting the equine piroplasmosis. The using method of the kit comprises the following steps:extracting DNA (deoxyribonucleic acid) from blood of a horse to be detected, further using the piroplasmosis universal primer to perform amplification, combining an amplification product with the detection film, enabling biotin at the terminal of a PCR (polymerase chain reaction) product to be in action with streptavidin-peroxidase, and then playing a catalysis role in enabling a substrate of peroxidase, namely an enhanced chemiluminescence reagent (ECL-detection) to emit fluorescence, thereby exposing an X-photosensitive film; and performing development and fixing treatment on the film so asto emerge a shadow in a region corresponding to specificity hybridization of a probe on the detection film and the PCR product, and further judging whether the detected animal is infected with the piroplasmosis or not according to whether the shade is left on the film or not.

Description

technical field [0001] The invention relates to a molecular biology identification and detection technology of animal parasites, specifically, the invention provides a detection kit for differential diagnosis of whether equine animals are infected with piriformis and a preparation and use method thereof. Background technique [0002] Equine piroplasmosis is an acute, subacute or chronic protozoal parasite caused by Babesia solani and Theileria equi parasitic in the red blood cells of equines (horses, donkeys, mules and zebras). The disease, which is transmitted by ticks, is mainly characterized by causing animal fever (sometimes periodically), anemia, jaundice, and hepatosplenomegaly. In the later stage of the disease, animals often have symptoms such as bilirubinuria and hemoglobinuria. Babesia caballi Nutall et Strickland, 1910 (old name horse coke, proplasma caballi) is a sub-kingdom of Protista (Protozoa), Apicomplexa, Sporozoea (Sporozoea) A member of the genus Babesia...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 刘光远谢俊仁田占成
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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