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Diagnostic kit and detection method of sheep Xinjiang babesia U sp.

A diagnostic kit and the technology of Babesia, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of chronic case limitations, reduce interference, reduce detection time, and improve accuracy Effect

Active Publication Date: 2017-01-25
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] For the diagnosis of Babesia, traditional methods are based on morphological examination of blood smears and clinical symptoms. These methods are very reliable for detecting acute cases, but have limitations for chronic cases.
Some serological tests are also commonly used for the diagnosis of Babesia, such as IFAT and ELISA, but these methods often cross-react with closely related species

Method used

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  • Diagnostic kit and detection method of sheep Xinjiang babesia U sp.
  • Diagnostic kit and detection method of sheep Xinjiang babesia U sp.
  • Diagnostic kit and detection method of sheep Xinjiang babesia U sp.

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Effect test

Embodiment 1

[0037] Example 1 Design and synthesis of specific primers:

[0038] According to the reported rap-1 gene sequence of Babesia ovis, after sequence comparison, specific primers for this species were designed and named in the conserved region of Babesia ovis xinjiang and the region that can be distinguished from other species For: XJ-fullF, XJ-fullR, XJ-internalF and XJ-internalR, dissolve the synthesized primers in deionized water to a storage concentration of 100uM, and find the optimal concentration for PCR amplification. The specific method is as follows: the primer is synthesized by a professional company according to the design requirements. After the conserved region is designed, the sequence of the primer is sent to the professional company to be synthesized by them. In the present invention, the primer is synthesized by Lanzhou Sangong Company. The specific primers for Babesia xinjiang were diluted with deionized water for use.

Embodiment 2

[0039] The preparation of embodiment 2 detection kits

[0040] 1. Preparation of Babesia xinjiang merozoites in sheep

[0041] Select 4 desplenic sheep for the test, and inoculate the jugular vein with blood red blood cells containing Theileria revneri, Theileria yoshii, Babesia moseni and Babesia xinjiang undetermined species preserved in liquid nitrogen, and infect the desplenic sheep.

[0042] After the sheep are infected, inject dexamethasone intramuscularly every day, 4 tubes / sheep for the first time, and 2 tubes / sheep thereafter, and inject once a day until the infection rate increases. From the date of infection, the body temperature of the sheep was measured every morning, and from the day when the body temperature began to rise, a blood smear was drawn from the vein of the ear edge every day, fixed with methanol twice, stained with Gimusa, and the infection rate of red blood cells was checked with a microscope. When the red blood cell infection rate reached above 10....

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Abstract

The invention discloses a diagnostic kit and a detection method for detecting sheep Xinjiang babesia U sp. through rapid nested PCR. The sheep Xinjiang babesia red blood cell invasion related protein (rap-1) is adopted as the target gene, and 4 bidirectional nested specific primers are designed and synthesized according to the rap-1 gene sequence to form specific primers XJ-full F(5'-CGGTAGTGTGTTGATATCCTAGAG-3'), XJ-full R(5'-CACAAGCGTTACATCGTAGAGCCG-3'), XJ-internal F(5'-AATGACCGCCCGCGAAATCAGCCACGA C-3') and XJ-internal R(5'-GTGGGTGGCGAAGGACAATTTGTTG-3'); and a nested PCR detection method is established for the sheep Xinjiang babesia of a field blood sample.

Description

technical field [0001] The invention relates to a diagnostic kit and a detection method for unspecified species of Babesia xinjiang, in particular to a diagnostic kit and a detection method for detecting unspecified species of Babesia xinjiang by using Nested-PCR. Background technique [0002] Babesiosis in sheep is a tick-borne blood protozoan disease of sheep and goats caused by Babesia spp., parasitic in the red blood cells of sheep or goats [1] . Babesia species that have been reported to infect sheep include Babesia ovis, B. motasi and B. crassa, which are mainly distributed in the tropics and subtropics. Blood protozoan diseases in the region. The clinical reaction is usually characterized by high fever, hemolytic anemia, jaundice, hemoglobinuria, and death in severe cases. The disease is serious, and can cause death in serious cases, causing significant economic losses to the sheep industry. Several Babesia isolates infecting sheep and goats were identified in Chi...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6848C12Q2549/119C12Q2521/301
Inventor 牛庆丽刘志杰杨吉飞潘玉萍关贵全罗建勋殷宏
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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