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Plasmodium detection method and detection system

A detection system and detection method technology, applied in the fields of biotechnology and medicine, can solve the problems of weak Raman signal and low detection sensitivity, etc.

Inactive Publication Date: 2018-07-27
SHENZHEN INT TRAVEL HEALTHCARE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main disadvantage of Raman laser detection of malarin is that the low content of malarin results in weak Raman signal, resulting in low detection sensitivity

Method used

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  • Plasmodium detection method and detection system
  • Plasmodium detection method and detection system
  • Plasmodium detection method and detection system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] With 10.4g / LRPMI 1640, 5.98g / LHEPES, 2g / L glucose, 0.025g / L hypoxanthine, 4.2g / LAlbumax II, 2.56g / L NaHCO 3 and 0.12g / L gentamicin-based complete medium, together with normal human fresh red blood cell suspension to cultivate Plasmodium, the Plasmodium strain comes from red blood cells infected with Plasmodium falciparum 3D7. After 3 days of culture, the culture medium was collected as a positive control sample.

[0087] 0.10g of saponin, 4g of NaCl, 0.1g of KCl, 1.8g of Na 2 HPO 4 ·12H 2 O, 0.12g of K 2 HPO 4 Dissolved in deionized water, made into a 500mL solution, stirred and mixed with a magnetic stirrer to obtain a lysis solution.

[0088] The positive control sample and the lysate were mixed at a volume ratio of 1:10 respectively. After fully lysed, centrifuged at 3000 rpm for 15 min to retain the first precipitate, and then repeated the operation of fully lysed, and centrifuged at 3000 rpm for 15 min to retain the second precipitate. The second pellet was w...

Embodiment 2

[0093] Normal human blood samples were used as negative control samples.

[0094] 0.10g of saponin, 4g of NaCl, 0.1g of KCl, 1.8g of Na 2 HPO 4 ·12H 2 O, 0.12g of K 2 HPO 4 Dissolved in deionized water, made into a 500mL solution, stirred and mixed with a magnetic stirrer to obtain a lysis solution.

[0095]Mix the negative control sample and the lysate at a volume ratio of 1:10, and centrifuge at 3000 rpm for 15 min after fully lysing to retain the first precipitate. Then repeat the operation of fully lysing and centrifuge at 3000 rpm for 15 min to retain the second precipitate. The second pellet was washed three times with saline and centrifuged at 3000 rpm for 15 min to retain the third pellet.

[0096] The third precipitate was placed in a detection dish, and the Raman signal was collected under the laser irradiation of a Raman spectrometer (LabRAM HR 800, HORIBA JobinYvon, France). Relevant experimental parameters of Raman spectrometer: 532nm wavelength laser genera...

Embodiment 3

[0101] 0.10g of saponin, 4g of NaCl, 0.1g of KCl, 1.8g of Na 2 HPO 4 ·12H 2 O, 0.12g of K 2 HPO 4 Dissolved in deionized water, made into a 500mL solution, stirred and mixed with a magnetic stirrer to obtain a lysis solution.

[0102] Dissolve 1.0 mL of frozen blood samples from patients with previous malaria infection detected by the medical laboratory of Shenzhen International Travel Health Care Center (confirmed as malaria infection by blood smear staining microscopy and PCR detection) and dissolve them in a volume ratio of 1:10. After fully lysing, centrifuge at 3000rpm for 15min to retain the first precipitate, then repeat the operation of fully lysing once, and centrifuge at 3000rpm for 15min to retain the second precipitate, then wash the second precipitate with physiological saline three times, and centrifuge at 3000rpm for 15min, retain the second precipitate. Three precipitation.

[0103] The third precipitate was placed in a detection dish, and the Raman signal...

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Abstract

The invention discloses a method for detecting malaria parasites, which comprises the following steps: providing a sample to be tested, and processing the sample to obtain a red blood cell suspension; preparing a saponin-containing lysate; mixing the red blood cell suspension with the lyse and fully After lysis, centrifuge and retain the first precipitate, then fully lyse the first precipitate, then centrifuge and retain the second precipitate, then wash the second precipitate with saline at least three times, then centrifuge and retain the third precipitate; use Raman laser irradiation The third step is to precipitate and collect the Raman signal; and analyze the Raman signal to determine whether the sample to be tested contains Plasmodium. This method of detection of malaria parasites releases malaria parasites from red blood cells through saponin, and then obtains malaria pigment by lysing the plasma parasite cell membrane, and concentrates malaria parasites. Combined with Raman spectroscopy, the detection sensitivity is high. The invention also discloses a malaria parasite detection system.

Description

technical field [0001] The invention relates to the fields of biotechnology and medicine, in particular to a detection method and a detection system for Plasmodium. Background technique [0002] Malaria is an insect-borne infectious disease caused by the infection of Plasmodium through the bite of Anopheles mosquitoes or the blood of people carrying Plasmodium. It is a disease that seriously endangers human health. Plasmodium is a black insoluble by-product produced by the digestion of hemoglobin in the host's red blood cells after Plasmodium infects humans, and can be used to determine the presence of Plasmodium infection. [0003] At present, the detection of malaria pigment is an important direction of malaria detection. It can not only be used to diagnose malaria infection, but also an important indicator to reflect the effect of clinical treatment. However, the current detection methods for malaria pigment have the shortcomings of weak detection signal and low sensitiv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/65G01N1/28
CPCY02A50/30
Inventor 董瑞玲顾大勇易品张树平朱玉兰刘胜牙
Owner SHENZHEN INT TRAVEL HEALTHCARE CENT
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