IL46963A

IL46963A - Reagent for detection of syphilis

Info

Publication number: IL46963A
Application number: IL46963A
Authority: IL
Original assignee: Centre Regional De Transfusion
Priority date: 1974-03-26
Filing date: 1975-03-28
Publication date: 1977-07-31
Also published as: JPS517119A; FR2266169B1; ES436045A1; FR2266169A1; JPS5315132B2; IL46963A0; SE7503428L; ZA751873B; DE2511688A1; GB1447470A; CH589294A5; CA1049403A

Abstract

A reagent useful in the detection of syphilis is disclosed. The reagent consists of an alcoholic solution of cardiolipids and is characterized by the fact that the lipids are stained in solution by means of a compatible dye. This reagent has the advantage that it can be kept and can be used in automatic blood analysis apparatus. [CA1049403A]

Description

FOR DETECTION OF SYPHILIS The present invention relates to a reagent which can be used to detect It is known that this disease can in fact be detected by a flocculation reaction between cardiolipids and blood serum or blood A of reagents are in fact which basically consist of an alcoholic solution of cardiolipids such as antigen or and by means of which syphilis be detected by performing laboratory reactions For it is a conventional practice to place a few drops of serum on a glass plate and then mix them with the antigen by If the reaction is positive the presence of a flocculate is detected Oh account of the development of automatic methods for performin various blood it became useful to include the detection of syphilis among the various systematic The automatic devices currently commercially available perform in parallel and five to seven analyses on serum depending on the detection of syphilis by automatic methods has raised some difficult problems for the investigators since the known antigens already mentioned the reactions carried out by hand are reagents which do not keep well and which have to be prepared shortly before the this preparation consists of diluting the alcoholic solution of cardiolipids in Of the apparatus known at the present time for carrying out these reactions those which work according to the principle may be which consists of introducing a sufficient amount of reagent into small quantities of serum or plasma from different which are separated by air All the samples of serum move continuously along flexible tubes of narrow with the aid of a The number tubes will be equal to the number of systematic analyses to be performed on the for example the blood group and syphilis After the reagents have been added to the samples the reaction products are poured onto absorbent thereby enabling the flocculation and thus the result to be observed a posteriori Such an apparatus available commercially under the trade mark TECHNICAN is described by McGrew in of a flocculation test for published by American Journal of Clinical In order to detect syphilis automatically it was thus necessary to have a reagent whose life is at least several weeks and which also produces very visible such that there will be no doubt about the or of the reactions whe they are for example when the reaction products are added to the a reagent which involves fixing the cardiolipids on particles of carbon has been developed instead of using an alcoholic solution of The blackish of these particles enables the test to be read very Details regarding the preparation and use of this reagent are given in Patent This which has been used by McGrew in automatic apparatus such as the TECHNICAN has given good this reagent has a serious disadvantage in that it is extremely In in certain cases it is found that the reaction is no longer visible on account of the fact that the antigen has been removed from the One important object of the reagent according to the invention is thus to detect syphilis since the life of reagent enables this to be performed and the reaction is made very visible when the flocculate is deposited on a sheet of absorben This reagent is much less expensive and is also extremely sensitive to the and it is possible to read the reaction very clearly and distinctly on account of the fact that the size of the lipid micelles in solution is The reagent according to the invention consists of an alcoholic solution of characterised by the fact that the lipids are stained in solution by means of a compatible The invention will be better understood on reading the following description of the reagent and also a process for the preparatio A dye for lipids which dissolves in alcohol and becomes on the lipids is added to the alcoholic solution of cardiolipids possibly comprising egg lecithin and maybe other more is added to commercially available or Various dyes have been tested and it can be said that black dyes give the best as least as regards the subsequent reading of the flocculation on the Amido Cerol black or Sudan black may be mentioned among the conventional dyes for Congo or ruthenium dyes are also The amount of dye used with respect to the alcoholic solution of cardiolipids is a function of the nature of the dye and its An amount of dye will be used which enables a saturated solution to be while avoiding an excess which might precipitate without Too small an amount of dye among other tend to reduce the sharpness of the Heating the mixture to for ten to forty minutes while shaking will stain the lipids by causing the dyes to be but will not denature the The mixture can then be diluted while hot with physiological In order to obtain a homogeneous formation of micelles during this dilution it is important not to cause any inuity in Physiological saline at the same erature as the between and will therefore be A preserving agent and various adjuvants for improving the sensitivity of the reaction can added to The automatic apparatus employed was a apparatus The syphilis detection was carried out by the Blood Transfusion Centre in order to determine whether the blood is free from disease and can therefore be used in Details the apparatus and its use in this field are given in the AMERICAN JOURNAL OF CLINICAL PATHOLOGY and in the bibliography The apparatus for employing the reagent in accordance with the invention was not A example of the preparation which has been found to be completely satisfactory in the automatic detection of both as regards the preservation of the product and the sharpness of the is given hereinafter 5 mg of Sudan black were added to 5 ml of a solution of The mixture was heated twenty minutes at about while The mixture was allowed to stand for and one of Sumatra benzoin was The stained kept at is then poured drop drop into 300 ml of physiological saline also heated to The micelles of the stained lipids which are obtained are homogeneous in ml of are then The ammonium sulphate increases the sensitivity of the The mixture is left to stand at ambient temperature and the reagent is then kept at for a g of choline chloride per 1 ml of solution is added to reduce the percentage of false positives positive The reagent may be kept for six weeks at Reagents of the same quality have also been prepared from antigen or the dyes already A reagent and a process for the preparation which is particularly suitable for detecting syphilis has been described and the mixture described above has bee found to be satisfactory when used in a continuous flow It will of course be possible for a person skilled in the art to produce minor depending on the desired eristics of the functioning of the apparatus by adding in order to obtain these For it would be possible to add to the reagent products for increasing the density of the micelles and their with a view to using the reagent in other types of in order to check the effectiveness and reliability of the reagent prepared in the example above the following procedure is adopted 389 samples of fresh serum were all of which gave four positive reactions using the following known methods Treponerae and Kolmer Cardiolipids Kline and insufficientOCRQuality