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Kits for the detection of Mycoplasma genitalium

A kit and mycoplasma technology, applied in the field of kits for detecting mycoplasma genitalium, can solve the problems of complex operation, uneven clinical performance, high cost of PCR, etc., achieve high sensitivity and specificity, easy popularization, and reduce economic burden Effect

Active Publication Date: 2022-04-08
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, PCR is expensive, complicated to operate, and its clinical performance is uneven, so it has not been accepted by clinicians and patients.

Method used

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  • Kits for the detection of Mycoplasma genitalium
  • Kits for the detection of Mycoplasma genitalium
  • Kits for the detection of Mycoplasma genitalium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Preparation of Mycoplasma genitalium detection card (Scheme 1)

[0016] 1. Weigh L-methionine p-nitroaniline, and use 0.2M NaH 2 PO 4 -Na 2 HPO 4 The buffer solution (pH4.0) was dissolved to make the concentration reach 0.2mg / ml, and mixed evenly to obtain the methionine aminopeptidase detection reagent. Add 10 ul / sheet of the liquid reagent to a circular paper sheet with a diameter of 6.0 mm, and dry to obtain the methionine aminopeptidase immobilized detection test paper.

[0017] 2. Weigh N-p-toluenesulfonyl-L-alanyloxynaphthylamine, with Na 2 HPO 4 -Dissolve the citric acid buffer (pH4.8) to make the concentration reach 0.5 mg / ml, mix well, and obtain the leukocyte esterase detection reagent.

[0018] Add 10ul / sheet of the liquid reagent to a circular paper sheet with a diameter of 6.0mm, and dry it to obtain the leukocyte esterase-immobilized test paper.

[0019] Fix the above two kinds of test papers in the flat shell-shaped test card in turn, and...

Embodiment 2

[0023] Example 2 The detection method of the Mycoplasma genitalium detection card prepared in Example 1

[0024] After diluting the genital tract secretion specimen with the treatment solution, take one drop (25~50ul) and add it on the paper for detecting methionine aminopeptidase and leukocyte esterase, and keep it under the condition of 37℃ for 15 minutes. Add one drop (25-40ul) of stop solution to the acid aminopeptidase disc, and read the enzyme activity results against the color comparison card within 1-3 minutes.

[0025]

[0026] Conclusion: When the results of methionine aminopeptidase and leukocyte esterase are positive, it indicates Mycoplasma genitalium infection.

Embodiment 3

[0027] Example 3 Preparation of Mycoplasma genitalium detection card (Protocol 2)

[0028] 1. Weigh L-proline p-nitroaniline, dissolve it with 0.4M Tris-citrate buffer (pH8.5) to make the concentration reach 1.5mg / ml, mix well, and obtain proline aminopeptidase detection reagent. Add 10ul / sheet of the liquid reagent to a circular paper sheet with a diameter of 6.0mm, and dry to obtain the proline aminopeptidase immobilized detection test paper.

[0029] 2. Weigh L-methionine p-nitroaniline, and use 0.2M NaH 2 PO 4 -Na 2 HPO 4 The buffer solution (pH4.0) was dissolved to make the concentration reach 0.2mg / ml, and mixed evenly to obtain the methionine aminopeptidase detection reagent. Add 10 ul / sheet of the liquid reagent to a circular paper sheet with a diameter of 6.0 mm, and dry to obtain the methionine aminopeptidase immobilized detection test paper.

[0030] 3. Weigh N-p-toluenesulfonyl-L-alanyloxynaphthylamine, use Na 2 HPO 4 -Dissolve the citric acid buffer (pH4.8...

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Abstract

The invention discloses a kit for detecting Mycoplasma genitalium, including two schemes: scheme one is two reagents, including methionine aminopeptidase and leukocyte esterase, wherein the activity of methionine aminopeptidase is detected The enzyme substrate used is a methionine derivative containing chromogen; the enzyme substrate used for detecting leukocyte esterase activity is an ester derivative containing chromogen. The second scheme is three kinds of reagents, that is, on the basis of the first scheme, it also includes a proline aminopeptidase detection reagent, and the enzyme substrate used for detecting the proline aminopeptidase activity is a proline derivative containing a chromogen. The invention detects the enzymes synthesized by the mycoplasma genitalium itself or the enzymes produced by stimulating parasitic cells through the dry chemical enzymatic method to achieve the purpose of detecting the mycoplasma genitalium, and has the advantages of quickness, simplicity, accuracy and the like. Using the kit of the invention to detect mycoplasma genitalium has high sensitivity and specificity, is similar in performance to existing similar detection reagents, and can fully meet the needs of clinical detection.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a kit for detecting mycoplasma genitalium. Background technique [0002] Mycoplasma genitalium (Mycoplasrna genitalium, Mg) belongs to the molluscum class Mycoplasma, is a prokaryote between bacteria and viruses, was first isolated by Tully in 1981 from 13 male patients with nongonococcal urethritis, It is the 12th species of mycoplasma discovered by humans. However, because the nutritional conditions of Mycoplasma genitalium are too harsh, it is extremely difficult to cultivate, and the culture period is long (greater than 50 days), few people conduct further research on it. Until 1985, Mullis K.B. et al., the human research laboratory of Getus Company in the United States, pioneered the DNA amplification technology of polymerase chain reaction (PCR). Using PCR technology, people conducted in-depth research on the pathogenicity of Mycoplasma genitalium and confirmed The imp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12Q1/37C12Q1/44C12R1/35
CPCC12Q1/04C12Q1/37C12Q1/44
Inventor 王则宇杨红云杨森刘志磊杨静静徐尚汪礼琴胡玉郭梦琳秦磊孙武举付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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