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Sputasol for isolated culture of Legionnella

A technology for separating and culturing and digesting liquid, applied in bacteria, biochemical equipment and methods, microorganisms, etc., can solve problems such as inability to digest, achieve the effect of improving the positive rate, facilitating clinical application, and reducing the pollution of miscellaneous bacteria

Inactive Publication Date: 2011-10-26
GUANGZHOU KINGMED DIAGNOSTICS CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the sputum digestive juice contains Na+ ions that inhibit the growth of Legionella, it cannot be used for digestion of sputum specimens isolated and cultured for Legionella
Therefore, there is still no sputum digestive fluid suitable for the isolation and cultivation of Legionella at home and abroad.

Method used

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  • Sputasol for isolated culture of Legionnella
  • Sputasol for isolated culture of Legionnella

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] 1. Preparation of stock solution (DTT~PBS 100ml): as shown in Table 1, weigh 2.0g of dithiothreitol (DTT), 0.894g of ethylenediaminetetraacetic acid (EDTA), 0.4g of potassium chloride (KCL), phosphoric acid Dipotassium Hydrogen (K 2 HPO 4 ) 3.6g, potassium dihydrogen phosphate (KH 2 PO 4 )0.4g add water (H 2 O) 100mL, mixed and dissolved, adjusted to pH 7.4, divided into vials, 5mL per bottle, and kept at room temperature for 6 months.

[0012] Table 1. Legionella isolation and culture sputum digestive fluid (stock solution 100ml volume)

[0013] Element

Content (g)

Dithiothreitol (DTT)

2.0

Ethylene Diamine Tetraacetic Acid (EDTA)

0.894

Potassium chloride (KCl)

0.4

Dipotassium hydrogen phosphate (Dipotassium hydrogen phosphate, K 2 HPO 4 )

3.6

Potassium dihydrogen phosphate (KH 2 PO 4 )

0.4

water (H 2 O)

100

[0014] 2. Preparation of working solution (0.1% DTT spu...

Embodiment 2

[0017] 1. Preparation of stock solution (DTT~PBS 100ml): weigh 1.5g of dithiothreitol (DTT), 0.892g of ethylenediaminetetraacetic acid (EDTA), 0.5g of potassium chloride (KCL), hydrogen phosphate Dipotassium (K 2 HPO 4 )3.2g, potassium dihydrogen phosphate (KH 2 PO 4 )0.8g add water (H 2 O) 100mL, mixed and dissolved, adjusted to pH 7.4, divided into vials, 4mL per bottle, and kept at room temperature for 6 months.

[0018] Table 2. Legionella isolation and culture sputum digestive fluid (stock solution 100ml volume)

[0019] Element

Content (g)

Dithiothreitol (DTT)

1.5

Ethylene Diamine Tetraacetic Acid (EDTA)

0.892

Potassium chloride (KCl)

0.5

Dipotassium hydrogen phosphate (Dipotassium hydrogen phosphate, K 2 HPO 4 )

3.2

Potassium dihydrogen phosphate (KH 2 PO 4 )

0.8

water (H 2 O)

100

[0020] 2. Preparation of working solution (0.1% DTT sputum digestion solution): Take 1 pa...

Embodiment 3

[0023] 1. Preparation of stock solution (DTT~PBS 100ml): as shown in Table 3, weigh 3.0g of dithiothreitol (DTT), 0.896g of ethylenediaminetetraacetic acid (EDTA), 0.3g of potassium chloride (KCL), hydrogen phosphate Dipotassium (K 2 HPO 4 ) 4.0g, potassium dihydrogen phosphate (KH 2 PO 4 ) 0.2g, add 100mL of water (H2O), mix and dissolve, adjust the pH to 7.4, divide into small bottles, 6mL per bottle, and keep at room temperature for 6 months.

[0024] Table 3. Legionella isolation and culture sputum digestive fluid (stock solution 100ml volume)

[0025] Element

Content (g)

Dithiothreitol (DTT)

3.0

Ethylene Diamine Tetraacetic Acid (EDTA)

0.896

Potassium chloride (KCl)

0.3

Dipotassium hydrogen phosphate (Dipotassium hydrogen phosphate, K 2 HPO 4 )

4.0

Potassium dihydrogen phosphate (KH 2 PO 4 )

0.2

water (H 2 O)

100

[0026] 2. Preparation of working solution (0.1% DTT sputum ...

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Abstract

The invention discloses a novel sputasol for isolated culture of Legionnella, which can be used for sputamentum digestion in isolated culture of Legionnella and can be also used as a digestive juice for conventional bacterial culture of clinical sputamentum. The sputasol comprises the following components: 2.0g of dithiothreitol (DTT), 0.894g of ethylene diamine tetraacetic acid (EDTA), 0.4g of potassium chloride, 3.6g of dipotassium hydrogen phosphate, 0.4g of potassium dihydrogen phosphate and 100g of water. By using the DTT, the sputasol breaks disulfide bonds in mucoproteins, thereby degrading the mucoproteins and digesting the sputamentum. By adding the EDTA into the sputasol, the EDTA can react with alkali metal, rare earth elements, transition metal and the like to form stable water-soluble complexes, thereby avoiding the poison of heavy metallic salts on the Legionnella. A buffer system comprises KCl, K2HPO4, KH2PO4 and other potassium salts and avoids the inhibiting effect of Na<+> ions on the Legionnella, thereby improving the positive rate in isolated culture of the Legionnella.

Description

technical field [0001] The invention relates to a sputum digestive liquid, in particular to a sputum digestive liquid suitable for the isolation and culture of Legionella in clinical sputum specimens. Background technique [0002] Legionella is an important pathogen that causes Legionnaires' disease, especially Legionella pneumophila infection, which is the main pathogen that causes Legionella pneumonia. For the diagnosis of Legionnaires' disease, bacterial isolation and culture is the gold standard. At present, the sputum digestive juice used for the isolation and culture of bacteria from clinical sputum specimens at home and abroad is Sputasol (Liquid) SR0233 produced by the British OXOID company. This digestive solution is only suitable for routine bacterial culture of clinical sputum specimens, not for Legionella culture. Because the phlegm digestive juice contains Na+ ions that inhibit the growth of Legionella, it cannot be used for the digestion of the sputum specimen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
Inventor 朱庆义胡朝晖赵利伟刘洋敏陈文聪
Owner GUANGZHOU KINGMED DIAGNOSTICS CENT
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