Leishmania liquid nutrient medium and preparation method thereof

A liquid culture medium, Leishmania technology, applied in microorganism-based methods, biochemical equipment and methods, protozoa, etc., can solve the problem of long time for detection results, low detection rate of Leishmania, and short validity period and other problems, to achieve the effect of long validity period, quick results and convenient storage.

Active Publication Date: 2013-11-13
新疆维吾尔自治区实验动物研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The invention provides a Leishmania liquid culture medium and a preparation method thereof, which overcomes the deficiencies of the above-mentioned prior art, and can effectively solve the problem of low detection rate and poor detection results of using 3N medium to culture and isolate Leishmania. The problem of long time and short validity period

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1: This Leishmania liquid culture medium contains NaCl 8.5g, KCl 0.4g, NaHCO 3 1.6g, KH 2 PO 3 0.2g, CaCl 2 0.2g, glucose 5.0g, L-glutamine 0.1g, Hepes 4.5g, RPMI-1640 10.4g, calf serum 200ml, deionized water 1700ml, penicillin 1 million units, streptomycin 1 million units, rabbit lysate Blood 100 ml; the Leishmania liquid culture medium is obtained according to the following steps: the first step, the above-mentioned required amount of NaCl, KCl, NaHCO 3 、KH 2 PO 3 , CaCl 2 , glucose and L-glutamine were dissolved and mixed in the above-mentioned required amount of deionized water, and the pH was adjusted to 7.2 with 0.1Mol / L NaOH solution, and the above-mentioned required amount of Hepes, RPMI-1640 and calf serum were added to mix In the second step, insert the inflatable tube into the lower part of the liquid, and fill the CO with the inflatable tube 2 gas into the liquid until the color of the liquid changes from light red to bright yellow, at w...

Embodiment 2

[0014] Embodiment 2: the difference with embodiment 1: the rabbit hemolysis of embodiment 2 obtains according to the following steps: the first step, selects the female rabbit of 1.3Kg to 1.5Kg, aseptic blood collection, glass bead method defibrillation, Add deionized water twice the volume of the blood after defibration, freeze and thaw repeatedly at a temperature between -5°C and 5°C until the red blood cells rupture, then centrifuge at 4000g for 20 minutes in a centrifuge, discard the precipitate, and the supernatant is ready Hemolysis for rabbits.

[0015] The above-mentioned embodiment gained Leishmania liquid culture medium is applied according to the following steps:

[0016] In the following practical applications, strictly follow the biosafety operation regulations.

[0017] 1 Preparation of equipment and materials for culturing and isolating Leishmania

[0018] 1. Equipment and equipment: biological safety cabinet, inverted microscope, cell culture bottle, capillar...

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Abstract

The invention relates to the technical field of liquid culture media for visceral leishmaniasis epidemic situation monitoring and visceral leishmaniasis diagnosis and discloses a leishmania liquid culture medium and a preparation method thereof. Compared with a traditional 3N culture medium, the leishmania liquid culture medium used for culturing suspected marrows and enlarged animal spleens of patients suffering from visceral leishmaniasis and culturing and separating leishmania has the advantages that separation ratio in culture and separation of the desert type visceral leishmaniasis is increased from less than 1% to higher than 10%; results are obtained rapidly, and result obtaining time after culture and observation is shortened from 15 days to 5 days; and convenience in preservation is achieved, and period of validity is prolonged from 30 days to 2 years. The leishmania liquid culture medium and the preparation method thereof provide bases for visceral leishmaniasis clinical diagnosis, epidemiological investigation and leishmania drug resistance study.

Description

technical field [0001] The invention relates to the technical field of liquid culture medium for kala-azar epidemic monitoring and kala-azar diagnosis, and relates to a Leishmania liquid culture medium and a preparation method thereof. Background technique [0002] Kala-azar is one of the Class B infectious diseases stipulated in my country's "Infectious Diseases Law". It is a natural focal parasitic disease caused by Leishmania. Sandflies are the medium of transmission, and sick people and animals are the main source of infection. ; The source of infection and the medium of transmission in some places have not yet been identified. According to the WHO report in 2012, leishmaniasis, including kala-azar, is widely distributed in 88 countries and regions, with 2 million new cases each year, and the total number of cases has reached 120 million. 350 million people are at risk of the disease. In recent years, the number of kala-azar patients in the world has been on the rise. Ka...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/10C12R1/90
Inventor 廖力夫徐艺玫燕顺生徐兵
Owner 新疆维吾尔自治区实验动物研究中心
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