JP2004141150A5

JP2004141150A5 -

Info

Publication number: JP2004141150A5
Application number: JP2003331963A
Authority: JP
Filing date: 2003-09-24
Publication date: 2006-11-09
Anticipated expiration: 2023-09-24

Claims

The red blood cells in the specimen are hemolyzed to release the malaria parasite, the free malaria parasite is stained with a fluorescent dye, a measurement sample is prepared, optical information is detected from the measurement sample, and the obtained optical information A malaria parasite detection method for distinguishing malaria parasites based on
The specimen is treated with a first hemolytic agent containing a first surfactant, and the treated specimen contains a second surfactant having a second hemolytic power stronger than that of the first surfactant. A method for detecting a malaria parasite, which is performed by treating with a hemolytic agent.

The second hemolytic agent sufficiently hemolyzes reticulocytes remaining in the specimen after being treated with the first hemolytic agent, and does not cause damage to the malaria parasite in the sample that affects the measurement. The method of claim 1 comprising:

The method of claim 1, wherein the first surfactant is selected from among nonionic surfactants and the second surfactant is selected from among cationic surfactants.

A first hemolytic agent containing a first surfactant, a second hemolytic agent containing a second surfactant having a hemolytic power stronger than that of the first surfactant, and a fluorescent dye A reagent kit for detecting malaria parasites.

The fluorescent dye is contained in either or both of the first hemolytic agent and the second hemolytic agent, or the fluorescent dye is packaged separately from the first hemolytic agent and the second hemolytic agent. The reagent kit according to claim 4.

The reagent kit according to claim 4 or 5, wherein the first surfactant is selected from among nonionic surfactants, and the second surfactant is selected from among cationic surfactants.

The nonionic surfactant has the following general formula:
_{_{_{R 1 -R 2 - (CH 2}}} CH 2 O) n-H
Wherein R ₁ is an alkyl, alkenyl, alkynyl group having 10 to 22 carbon atoms, R ₂ is —O—, — (C ₆ H ₄ ) —O— or —COO—, and n is an integer of 8 to 120. is there)
The reagent kit of Claim 6 represented by these.

Cationic surfactant has the following general formula

(Wherein R ^a is an alkyl, alkenyl or alkynyl group having 8 to 20 carbon atoms, R ^b , R ^c and R ^d are alkyl, alkenyl or alkynyl groups having 1 to 8 carbon atoms, and X − and Y − are anions. , M is an integer from 7 to 19)
The reagent kit of Claim 6 represented by these.

The reagent kit according to claim 4 or 5, wherein the fluorescent dye is a fluorescent dye for staining nucleic acid.

Fluorescent dyes are in the following groups:

(Wherein R1 is a hydrogen atom or a lower alkyl group; R2 and R3 are a hydrogen atom, a lower alkyl group or a lower alkoxy group; R4 is a hydrogen atom, an acyl group or a lower alkyl group; R5 is a hydrogen atom or a substituted one) A good lower alkyl group; Z is a carbon atom substituted with a sulfur atom, an oxygen atom or a lower alkyl group; n is 1 or 2; X- is an anion),

Wherein R 1 is a hydrogen atom or an alkyl group; R 2 and R 3 are a hydrogen atom, a lower alkyl group or a lower alkoxy group; R 4 is a hydrogen atom, an acyl group or an alkyl group; Z is a sulfur, oxygen or lower alkyl group Substituted carbon atoms; n is 0, 1 or 2; X- is an anion), as well as acridyline orange, thiazole orange, ethidium bromide, propidium iodide, auramine O, Hoechst 33258, Hoechst 33342, The reagent kit according to claim 4 or 5, which is selected from Rhodamine 123 and DiOC1 (3).

The reagent kit according to claim 4 or 5, wherein the osmotic pressure of the first hemolytic agent and the second hemolytic agent is 150 to 600 mOsm / kg.

A sample containing malaria parasite is treated with a first hemolytic agent containing a first surfactant, and the sample treated with the first hemolytic agent is treated with a second interface having a stronger hemolytic force than the first surfactant. A chamber containing a measurement sample obtained by treating with a second hemolytic agent containing an active agent and staining the sample treated with the second hemolytic agent with a fluorescent dye;
A flow cytometer for measuring fluorescence information of the measurement sample, a control unit for discriminating malaria parasites based on the fluorescence information,
A malaria parasite detector.

The detection apparatus according to claim 12, wherein a flow cytometer measures fluorescence information of a measurement sample, and a control unit discriminates malaria parasites based on the fluorescence information and scattered light information.