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Method and device for establishment of Mycobacterium tuberculosis naturally infected non-human primate model

A primate and non-human technology, applied in animal houses, medical raw materials derived from bacteria, animal husbandry, etc., can solve the problems that cannot be simulated as latent infection

Inactive Publication Date: 2014-11-05
WUHAN UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0020] For the existing TB macaque models, Mycobacterium tuberculosis is directly inoculated into the trachea or lung of macaques in an artificial way (such as fiberoptic bronchoscopy or bronchial inoculation), and most (60-80%) artificially infected macaques develop acute activity Sexual tuberculosis, rapid death in a short period of time, can not simulate the defects of most of the processes of latent infection after human infection with Mycobacterium tuberculosis, the invention provides a method to establish natural M.tb infection (airborne transmission) ) method and device for a non-human primate model

Method used

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  • Method and device for establishment of Mycobacterium tuberculosis naturally infected non-human primate model
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  • Method and device for establishment of Mycobacterium tuberculosis naturally infected non-human primate model

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preparation example Construction

[0055] 1. Preparation of Mycobacterium tuberculosis

[0056] The strain used to infect animals is Mycobacterium tuberculosis H37Rv strain, take 1×10 7 CFU can cause half of the infected mice to die in about 2 weeks (determined by pre-testing) Mycobacterium tuberculosis H37Rv strain preserved on a slant, use a sterile inoculation loop to scrape a little bacteria lawn on the slant, inoculate it on a freshly improved On the slant of Roche's medium, culture at 37°C for 3 weeks. Using a sterile culture ring, scrape the bacterial lawn on the inclined surface, weigh it accurately and grind it in a sterile mortar, add sterile saline at 10-20 mg / mL, and grind it while adding until it is completely dispersed and uniform Cloudy bacteria. Use a quantitative sterile pipette to distribute the bacterial suspension into 1.5mL cryopreservation tubes, 1mL per tube, put them into a marked freezer box, and store them in a -80°C refrigerator as a strain preservation tube. .

[0057] The bacter...

Embodiment 1

[0092] [Example 1] Fiberoptic bronchoscopic infection of Chinese macaques

[0093] 1.1 Anesthesia

[0094] 2 experimental monkeys WNP1 and WNP2 were fasted for 8-12 hours before anesthesia. 15 minutes before anesthesia, intramuscular injection of atropine 0.04 mg / kg body weight and intramuscular injection of ketamine hydrochloride 10 mg / kg body weight gave anesthesia. Afterwards, the animals were placed on the operating table.

[0095] 1.2 Animal infection

[0096] After the animal is anesthetized, lie supine on the operating table, open the mouth with a gag, expose the epiglottis with a laryngoscope, and anesthetize the epiglottis with 1% tetracaine. Operate according to the instructions of the fiber bronchoscope (OLYMPUS BF TYPE XP60, outer diameter 2.8mm), spray 1% lidocaine into the nostrils on both sides of the animal with a sprayer, slowly insert the fiber bronchoscope through the nasal passage of the animal, and when it reaches the epiglottis , slowly inject 1mL of 1...

Embodiment 2

[0099] [Example 2] Mycobacterium tuberculosis infects Chinese rhesus monkeys by natural route—aerosol

[0100] WPN1 and WPN2 rhesus monkeys that were artificially infected (direct inoculation of Mtb strain into the lungs by fiberoptic bronchoscopy) were placed in an airflow self-circulation combined negative pressure isolator on the day after the artificial infection (see Figure 7 ), and then put 4 healthy monkeys WPN3, WPN4, WPN5, WPN6 into the combined negative pressure isolator, start the airflow self-circulation system, so that the healthy monkeys can repeatedly contact the air from the infected monkey cage, and fresh air can be supplied as required , Control the air supply and exhaust through the PLC control unit, and adjust the air volume. After exposure, clinical symptoms such as appetite and cough were observed every day; various tests were arranged according to Table 3.

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Abstract

The invention provides a method for establishment of a Mycobacterium tuberculosis naturally infected non-human primate model. According to the invention, a healthy non-human primate and a non-human primate artificially infected with tuberculosis and suffering cough are raised in an air flow self-circulation combined type negative pressure isolator so as to infect the healthy non-human primate with tuberculosis. Establishment of the natural infection model helps to reveal conditional factors (like HIV) activating latent tuberculosis infection and related mechanisms. The non-human primate model with more clinical significance is provided for research, development and evaluation of anti-tuberculosis vaccines and drugs.

Description

technical field [0001] The invention relates to a method and a device for establishing an animal model, in particular to a method and a device for establishing a model of a non-human primate naturally infected by Mycobacterium tuberculosis. Background technique [0002] Tuberculosis (TB) is one of the most common and refractory respiratory infectious diseases worldwide caused by Mycobacterium tuberculosis (M.tb). It is a global public health problem that seriously endangers public health [1]. Since the 1980s, due to the increase of floating population, the rapid spread of AIDS, the limitation of BCG vaccine preventive effect and the emergence of multi-drug resistant M. The mortality rate has shown an obvious upward trend, which has brought new and severe challenges to the prevention and control of tuberculosis. According to the World Health Organization report: 2 billion people worldwide are infected with M.tb. my country is the second largest country with the highest bur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/74A01K1/03A01K1/035A01K1/00
Inventor 霍文哲付博文郭铭李湘东王勇
Owner WUHAN UNIV
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