Direct fungus detection tachypleus amebocyte lysate box and method

The technology of a Limulus reagent and a kit is applied in the field of direct fungus detection Limulus kits, which can solve the problems of no fungi being detected, long time and the like.

Inactive Publication Date: 2010-11-10
丁友玲
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, blood culture is used clinically to diagnose fungal infection, which takes a relatively long time, and due to the extensive use of broad-spectrum antibiotics, many patients cannot detect fungi by blood culture

Method used

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  • Direct fungus detection tachypleus amebocyte lysate box and method
  • Direct fungus detection tachypleus amebocyte lysate box and method
  • Direct fungus detection tachypleus amebocyte lysate box and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] 1. Reagent preparation:

[0055]1) 0.05mol / L buffer solution (Tris-HCl / pH8.0): Weigh 6.057g of Tris base, first add 500ml of sterile pyrogen-free water to dissolve, then add 60ml of 37% concentrated hydrochloric acid to adjust the pH to about 8.0, and then set Capacity up to 1000ml.

[0056] 2) Washing solution: 2mol / LNaCl / 0.1mol / LTris-HCl / pH8.0, weigh 117g of sodium chloride after dry heat sterilization, and then dilute to 1000ml with 0.1mol / LTris-HCl / pH8.0.

[0057] 3) Disintegration solution: 0.1mol / LNaCl / 0.1mol / LTris-HCl / pH8.0, preparation: weigh 5.85g of sodium chloride after dry heat sterilization, then add 0.1mol / LTris-HCl / Ph8.0 Set the volume to 1000ml;

[0058] 4) Eluent:

[0059] ①0.03mol / LNaCl / 0.015mol / LTris-HCl / pH8.0

[0060] Weigh 1.755 g of sodium chloride after dry heat sterilization, and then use 0.015 mol / LTris-HCl / pH8.0 to adjust the volume to 1000 ml.

[0061] ②0.2mol / LNaCl / 0.015mol / LTris-HCl / pH7.4~8.2

[0062] Weigh 11.7 g of sodium chloride af...

Embodiment 2

[0104] Example 2: Clinical Evaluation of Direct Fungal Detection Limulus Kit (G Reagent)

[0105] 123 specimens were tested according to the method described in Example 1, and compared with the traditional fungal culture method. The results are shown in Table 3.

[0106] Table 3 Comparison of results of fungal detection methods

[0107]

[0108] Note: X2=400, P<0.05, the difference is significant

[0109] As shown in Table 3, the number of positive cases by the plasma dextran detection method is 81 cases, and these 81 cases are also detected by the traditional fungal culture method, and the number of positive cases is 73 cases, and another 8 cases are negative; The number of negative cases was 42 cases, and the results of these same traditional fungal culture methods showed that one case was positive and the other 41 cases were negative.

[0110] It can be seen from Table 3 that among the 123 specimens, 74 cases were positive for fungus cultured by traditional methods...

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Abstract

The invention discloses a direct fungus detection tachypleus amebocyte lysate box and a direct fungus detection method. The kit comprises: (1) a G reagent which is a mixture of lichaampje cytolyzate and color development material of tachypleus; (2) buffer solution which is 0.02 to 0.1mol / L Tris-HCL; (3) a standard substance which is (1-3)-beta-D-glucan; (4) diazotizing agent; and (5) blood treating liquid. The (1-3)-beta-D-glucan content of a sample is calculating by processing the sample and the standard substance, preparing solution of the G reagent, filling, raising temperature, stopping, measuring a light-absorbing value and making a standard curve. The kit can quickly and accurately measure the micro (1-3)-beta-D-glucan content of human body blood and body blood samples.

Description

technical field [0001] The present invention relates to a kit and its use method, more specifically to a kit and method for direct fungal detection of Limulus. Background technique [0002] At present, it is still a very difficult problem for clinicians to diagnose whether a patient has fungal infection, especially deep fungal infection or fungal sepsis. Currently, blood culture is used clinically to diagnose fungal infection, which takes a relatively long time, and due to the extensive use of broad-spectrum antibiotics, many patients cannot detect fungus by blood culture. In this context, clinicians urgently need a reagent that can quickly and accurately determine whether it is a fungal infection. Contents of the invention [0003] In order to solve the above problems, the present invention provides a direct fungal detection Limulus test kit and method, which can quickly and accurately quantify the trace amount of (1-3)-β-D-glucan in human blood and body fluid specime...

Claims

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Application Information

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IPC IPC(8): C12Q1/04G01N21/31
Inventor 丁友玲廖招连
Owner 丁友玲
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