SE431228B - SETTING FOR DETERMINATION OF BACTERIAL ENDOTOXIN MEDIUM PROPHENOLOXIDAS-CONTAINING ANIMAL ANIMAL OR INSECTS AND REAGENTS - Google Patents

SETTING FOR DETERMINATION OF BACTERIAL ENDOTOXIN MEDIUM PROPHENOLOXIDAS-CONTAINING ANIMAL ANIMAL OR INSECTS AND REAGENTS

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Publication number
SE431228B
SE431228B SE8107599A SE8107599A SE431228B SE 431228 B SE431228 B SE 431228B SE 8107599 A SE8107599 A SE 8107599A SE 8107599 A SE8107599 A SE 8107599A SE 431228 B SE431228 B SE 431228B
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SE
Sweden
Prior art keywords
compound
insects
animal
oxidase
lysate
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SE8107599A
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Swedish (sv)
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SE8107599L (en
Inventor
Kenneth Soderhell
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Kenneth Soderhell
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Publication date
Application filed by Kenneth Soderhell filed Critical Kenneth Soderhell
Priority to SE8107599A priority Critical patent/SE431228B/en
Priority to JP83500092A priority patent/JPS58502082A/en
Priority to EP83900080A priority patent/EP0096689A1/en
Priority to PCT/SE1982/000430 priority patent/WO1983002123A1/en
Publication of SE8107599L publication Critical patent/SE8107599L/en
Publication of SE431228B publication Critical patent/SE431228B/en

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/579—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving limulus lysate
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/0808—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2400/00—Assays, e.g. immunoassays or enzyme assays, involving carbohydrates
    • G01N2400/10—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • G01N2400/12—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar
    • G01N2400/24—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar beta-D-Glucans, i.e. having beta 1,n (n=3,4,6) linkages between saccharide units, e.g. xanthan

Description

l0 15 20 25 30 35 8107599-6 2 svampar. Även om den biokemiska mekanismen inte är helt klarlagd, så aktiverar lipopolysackariderna resp. ß-lß-glukanerna ett serinproteas, som förutom att omvandla koagulogen till koagulin, överför profenoloxidas till det aktiva enzymet fenoloxidas. Bildningen av fenoloxidas vid den senare aktiveringen kan enligt uppfinningen utnyttjas för att påverka en lämplig detektorförening till en förening som kan påvisas fysikaliskt eller kemiskt. l0 15 20 25 30 35 8107599-6 2 mushrooms. Although the biochemical mechanism is not fully understood, the lipopolysaccharides activate resp. The ß-lß-glucans are a serine protease which, in addition to converting the coagulogen to coagulin, converts profenol oxidase to the active enzyme phenol oxidase. The formation of phenol oxidase in the latter activation can according to the invention be used to effect a suitable detector compound into a compound which can be detected physically or chemically.

Enligt uppfinningen kan således en bakterieinfektion snabbt och enkelt påvisas genom att man för ett prov som skall undersökas i kontakt med dels ett buffrat profenoloxidashaltigt blodkroppslysat från någon av artropodklasserna kräftdjur eller insekter, och dels en detektorsubstans i form av åtminstone en fenolförening med förmåga att oxideras av enzymet fenoloxidas till en fysikaliskt eller kemiskt, t.ex. spektrofotometriskt eller kolorimetriskt, pâvisbar förening.Thus, according to the invention, a bacterial infection can be detected quickly and easily by passing a buffer to be examined in contact with a buffered propenol oxidase-containing blood cell lysate from one of the arthropod classes crustaceans or insects, and a detector substance in the form of at least one phenolic compound capable of oxidizing the enzyme phenol oxidase to a physical or chemical, e.g. spectrophotometric or colorimetric, detectable compound.

Ett motsvarande reagens eller reagenssats för påvisande av bakterieinfektioner innefattar därför dels ett sådant blodkroppslysat från kräftdjur eller insekter och dels en sådan detektorsubstans. _ Även om generellt sett blodkroppslysat från alla kräftdjur och insekter skulle kunna användas enligt uppfinningen, så är bland kräftdjuren (Crustacea) de tiofotade kräftdjuren eller de s.k. dekapoderna föredragna. Såväl sötvattens- dekapoder som marina sådana kan användas. Som exempel på sötvattenskräftor kan nämnas Astacus astacus (flodkräfta), Pacifastacus leniusculus (signalkräfta), Astacus leptodactylus, Cambarus affinis (nordamerikansk flodkräfta), Procam- barus clarkii. Bland lämpliga marina dekapoder kan nämnas Cancer pagurus (krabbtaska), Carcinusmaenas (strandkrabba), l-lomarus vulgaris (hummer), Palinurus vulgaris (langust), Nephrops norvegicus (kejsarhummer). Bland dessa kräftdjur är flera direkt lämpade för odling i akvakultur, t.ex. flodkräfta och signalkräíta, men även hummer och krabba, och är därför att föredra för uppfinningens syfte.A corresponding reagent or reagent kit for the detection of bacterial infections therefore comprises on the one hand such a blood cell lysate from crustaceans or insects and on the other hand such a detector substance. Although in general blood cell lysate from all crustaceans and insects could be used according to the invention, among the crustaceans (Crustacea) the ten-footed crustaceans or the so-called decapods preferred. Both freshwater decapods and marine ones can be used. Examples of freshwater crayfish are Astacus astacus (crayfish), Pacifastacus leniusculus (signal crayfish), Astacus leptodactylus, Cambarus affinis (North American crayfish), Procambarus clarkii. Suitable marine decapods include Cancer pagurus (crab bag), Carcinusmaenas (beach crab), l-lomarus vulgaris (lobster), Palinurus vulgaris (lobster), Nephrops norvegicus (emperor lobster). Among these crustaceans, several are directly suitable for cultivation in aquaculture, e.g. crayfish and signal crayfish, but also lobster and crab, and is therefore preferred for the purpose of the invention.

Bland insekterna kan särskilt nämnas sådana tillhörande Orthoptera och Lepídoptera (fjärilar). Exempel på den förstnämnda ordningen är Schistocerca gregaria (ökengräshoppa) och Locusta migratoria (sträckgräshoppa). Bland fjäri- larna kan nämnas Galleria melonella (vaxmott), l-lyalphora cecropia och Bombyx mori (silkesfjärll). Även om insekterna inte torde vara lika aktuella för lysatframställning som kräftdjuren, så skulle man mycket väl kunna tänka sig odling av exempelvis silkesfjärilar för detta ändamål.Among the insects, special mention may be made of those belonging to Orthoptera and Lepídoptera (butterflies). Examples of the first-mentioned order are Schistocerca gregaria (desert grasshopper) and Locusta migratoria (stretch grasshopper). The butterflies include Galleria melonella (wax moth), l-lyalphora cecropia and Bombyx mori (silkworm). Although the insects should not be as relevant for lysate production as the crustaceans, it would very well be conceivable to grow silk butterflies for this purpose, for example.

Lämpliga fenolföreningar för användning som detektorföreningar är t.ex. kinonföreningar med karakteristisk färg, särskilt dihydroxibensenföreningar eller föreningar, som innefattar en sådan dihydroxisubstituerad 'ienylgrupp. En speciellt lämplig förening är dihydroxifenylalanin (DOPA), som efter oxidation 10 15 20 25 30 35 8107599-6 med fenoloxidas ger en kraftigt röd färg vid 490 nm, som är stabil i ungefär 5 - 6 timmar. Denna substans finns kommersiellt tillgänglig till relativt låg kostnad och kan för uppfinningens ändamål användas i tämligen oren form. Ett annat exempel är ß-metylkatekol, som ger en lila färg vid 520 nm. Den bildade kinonen är dock relativt instabil och stabiliseras därför lämpligtvis genom närvaro av hydroxiprolinester.Suitable phenolic compounds for use as detector compounds are e.g. quinone compounds of characteristic color, especially dihydroxybenzene compounds or compounds comprising such a dihydroxy-substituted phenyl group. A particularly suitable compound is dihydroxyphenylalanine (DOPA), which after oxidation with phenol oxidase gives a strong red color at 490 nm, which is stable for about 5-6 hours. This substance is commercially available at a relatively low cost and can be used for the purposes of the invention in rather crude form. Another example is β-methyl catechol, which gives a purple color at 520 nm. However, the quinone formed is relatively unstable and is therefore conveniently stabilized by the presence of hydroxyproline ester.

Ett hemocytlysat, dvs. blodkroppslysat, från kräftdjur och insekter enligt uppfinningen kan framställas principiellt på samma sätt som det välkända limuluslysatet från hästskokrabban. Denna metod finns väl beskriven tidigare i litteraturen och behöver därför inte redovisas närmare här. Sålunda kan ett partiellt renat hemocytlysat från exempelvis kräftdjur framställas genom att man först tappar djuret på blod, varvid man ser till att förorening av blodet undviks. Det bör här framhållas, att ett kräftdjur, som kan odlas i akvakultur, som t.ex. flodkräfta, signalkräfta etc. inte behöver avlivas vid blodtappningen, utan att man kan tappa samma djur ett flertal gånger med jämna mellanrum i likhet med en mänsklig blodgivare. Detta är givetvis en stor fördel, eftersom då utvinning av blod för hemocytlysatframställning enligt uppfinningen kan kom- bineras med kräftodling för livsmedelsändamål. Därefter isoleras hemocyterna eller blodkropparna genom centrifugering och tvättning på konventionellt sätt.A hemocyte lysate, i.e. blood cell lysate, from crustaceans and insects according to the invention can be prepared in principle in the same way as the well-known limulus lysate from the horseshoe crab. This method is well described earlier in the literature and therefore does not need to be described in more detail here. Thus, a partially purified hemocyte lysate from, for example, crustaceans can be prepared by first draining the animal of blood, thereby ensuring that contamination of the blood is avoided. It should be emphasized here that a crustacean, which can be grown in aquaculture, such as crayfish, signal crayfish, etc. do not have to be killed during the blood draw, without being able to lose the same animal several times at regular intervals, similar to a human blood donor. This is of course a great advantage, since then the extraction of blood for hemocyte lysate production according to the invention can be combined with crayfish cultivation for food purposes. Thereafter, the hemocytes or blood cells are isolated by centrifugation and washing in a conventional manner.

Man homogeniserar sedan i buffert med hög kalciumjonkoncentration, centri- fugerar vid ca 70.000 g och tar till vara supernatanten. Det erhållna lysatet håller sig stabilt i ca l dygn. Företrädesvis frystorkas emellertid den erhållna supernatanten för att vid användningstillfället spädas med vatten eller lämplig buffert (prio/ß). Eventuellt kan lösningen stabiliseras med 0,5 - 1,5 NaCl, varigenom en stabilitet på flera dagar kan uppnås.It is then homogenized in a buffer with a high calcium ion concentration, centrifuged at about 70,000 g and the supernatant used. The lysate obtained remains stable for about 1 day. Preferably, however, the resulting supernatant is lyophilized to be diluted with water or a suitable buffer (prio / ß) at the time of use. Optionally, the solution can be stabilized with 0.5 - 1.5 NaCl, whereby a stability of several days can be achieved.

Ett reagens eller en reagenssats enligt uppfinningen för att påvisa bakterieinfektioner innehåller ett blodkroppslysat framställt enligt ovan och en fenolförening enligt den tidigare definitionen. Företrädesvis föreligger lysatet och detektorsubstansen i pulverform. Vid användning för att testa ett extrakt med misstänkt bakterieinfektion löser man testreagenset i lämplig buffert (pH/IX), varefter man tillsätter det extrakt som skall testas. Reagenslösningen studeras därefter exempelvis spektrofotometriskt eller kolorimetriskt beroende på den använda detektorsubstansen.A reagent or reagent kit according to the invention for detecting bacterial infections contains a blood cell lysate prepared as above and a phenolic compound as previously defined. Preferably, the lysate and the detector substance are in powder form. When used to test an extract with suspected bacterial infection, the test reagent is dissolved in a suitable buffer (pH / IX), after which the extract to be tested is added. The reagent solution is then studied, for example, spectrophotometrically or colorimetrically, depending on the detector substance used.

Uppfinningen kommer nu att beskrivas närmare med hjälp av några speciella utföringsexempel, som dock inte på något sätt är begränsande för uppfinningen. _5 10 15 20 25 8107599-6 4 Exempel l Framställning av hemocytlysat l-lemocyter från flodkräfta, Astacus astacus, uppsamlades såsom beskrivs i Söderhäll, K., Häll, L., Unestam, T., och Nyhlen, L. (1979) J. Invertebr. Pathol. fi, 285-294, med undantag av att 0,1 M natriumcitrat inte användes. Hemocy- terna homogeniserades i 10 mM natriumkakodylatbuffert, pH 7,0, med 100 mM CaClz, och homogenatet centrifugerades sedan 20 minuter vid 70.000 g. Den erhållna supernatanten som innehåller ungefär 2 mg protein/ml, kan användas antingen direkt eller frystorkas i alikvoter om 4 ml. För användning löses den i 2,9 ml destillerat vatten till en slutlig proteinkoncentration på 2 mg/ml.The invention will now be described in more detail with the aid of some special embodiments, which, however, are in no way limiting of the invention. Example 1 Preparation of Hemocyte lysate I-lemocytes from crayfish, Astacus astacus, were collected as described in Söderhäll, K., Häll, L., Unestam, T., and Nyhlen, L. (1979) J. Invertebr. Pathol. fi, 285-294, except that 0.1 M sodium citrate was not used. The hemocytes were homogenized in 10 mM sodium cacodylate buffer, pH 7.0, with 100 mM CaCl 2, and the homogenate was then centrifuged for 20 minutes at 70,000 g. The resulting supernatant containing about 2 mg protein / ml can be used either directly or lyophilized in aliquots if 4 ml. Before use, it is dissolved in 2.9 ml of distilled water to a final protein concentration of 2 mg / ml.

Exempel 2 Två volymer kräfthemocytlysat enligt Exempel 1 inkuberades med en volym Zymosan-supernatant (supernatanten från l96-ig suspension av jäst- cellväggar, Sigma) vid 20°C. Omedelbart efter tillsatsen bestämdes aktiviteten av serinproteas resp. fenoloxidas med jämna mellanrum under ca 60 minuter.Example 2 Two volumes of cancer hemocyte lysate of Example 1 were incubated with one volume of Zymosan supernatant (the supernatant from the 96-well suspension of yeast cell walls, Sigma) at 20 ° C. Immediately after the addition, the activity of serine protease resp. phenol oxidase at regular intervals for about 60 minutes.

Serinproteasaktiviteten analyserades genom att man inkuberade 100 pl av reaktionsblandningen, 600 pi 0,1 M-tris-HCl-buffert, pH 8,0, och 100 pl med den syntetiska peptiden Bz-Ile-Glu-(Y-O-piperidyD-Gly-Arg-PNA-l-ICI (AB Kabi Pep- tidforskning). Reaktionen avslutades efter inkubering i 20 minuter vid 37°C genom tillsats av 100 pl 5096-ig ättiksyra. Enzymaktiviteten visas i den bifogade figuren som förändringen i absorbans vid 1405 nm.Serine protease activity was assayed by incubating 100 μl of the reaction mixture, 600 μl of 0.1 M-Tris-HCl buffer, pH 8.0, and 100 μl of the synthetic peptide Bz-Ile-Glu- (YO-piperidyD-Gly-Arg PNA-1-ICI (AB Kabi Pep Time Research) The reaction was terminated after incubation for 20 minutes at 37 ° C by the addition of 100 μl of 5096 μg acetic acid.The enzyme activity is shown in the accompanying figure as the change in absorbance at 1405 nm.

Fenoloxidasaktiviteten bestämdes genom att man inkuberade 100 pl av reaktionsblandningen med 30 pl L-dopa (4 g/1) i 5 minuter vid 490 nm. Enzym- aktiviteten uttrycks i figuren som förändringen i absorbans vid 490 nm.Phenol oxidase activity was determined by incubating 100 μl of the reaction mixture with 30 μl of L-dopa (4 g / l) for 5 minutes at 490 nm. The enzyme activity is expressed in the figure as the change in absorbance at 490 nm.

"X-x" i Figuren avser kontroll med buffert (0,01 M natriumacetat, pH 5,2) stället för B-lß-glukantillsats."X-x" in the Figure refers to control with buffer (0.01 M sodium acetate, pH 5.2) instead of B-β1-glucan additive.

Ur figuren framgår klart den stabilare nfenoloxidasaktiviteten.The figure clearly shows the more stable nphenol oxidase activity.

Claims (8)

1. l0 15 20 25 30 8107599-6 5 PATENTKRAV l. Sätt att bestämma bakteriellt endotoxin, k ä n n e t e c k n a t av att man för det prov som skall undersökas i kontakt med (A) ett buffrat profenoloxidashaltigt blodkroppslysat från ett djur till- hörande någon av artropodklasserna kräftdjur (Crustacea) och insekter (lnsecta), och (B) en detektorsubstans i form av åtminstone en fenolförening med förmåga att oxideras av enzymet fenoloxidas till en förening som kan påvisas fysikaliskt eller kemiskt, t.ex. spektrofotometriskt eller kolorimetriskt.1. Method of determining bacterial endotoxin, characterized in that for the sample to be examined in contact with (A) a buffered propenol oxidase-containing blood cell lysate from an animal belonging to any of the the arthropod classes crustaceans (Crustacea) and insects (lnsecta), and (B) a detector substance in the form of at least one phenolic compound capable of being oxidized by the enzyme phenol oxidase to a compound which can be detected physically or chemically, e.g. spectrophotometric or colorimetric. 2. Sätt enligt patentkravet 1, k ä n n e t e c k n a t av att íenolföreningen är en dihydroxibensenförening.2. A method according to claim 1, characterized in that the phenolic compound is a dihydroxybenzene compound. 3. Sätt enligt patentkravet 2, k ä n n e t e c k n a t av att fenolföreningen är dihydroxifenylalanin eller 4- metylkatekol.3. A process according to claim 2, characterized in that the phenolic compound is dihydroxyphenylalanine or 4-methyl catechol. 4. Sätt enligt något av patentkraven 1-3, k ä n n e t e c k n a t av att detektorsubstansen innefattar ett stabiliserings- medel för den oxiderade föreningen.4. A method according to any one of claims 1-3, characterized in that the detector substance comprises a stabilizing agent for the oxidized compound. 5. Sätt enligt patentkravet 4, k ä n n e t e c k n a t av att fenolföreningen är lß-metylkatekol och att stabili- seringsmedlet är hydroxiprolinester.5. A process according to claim 4, characterized in that the phenolic compound is β-methyl catechol and that the stabilizing agent is hydroxyproline ester. 6. Sätt enligt något av patentkraven 1-5, k ä n n e t e c k n a t av att blodkroppslysatet är ett hemocytlysat från ett djur tillhörande ordningen dekapoder.6. A method according to any one of claims 1-5, characterized in that the blood cell lysate is a hemocyte lysate from an animal belonging to the order decapods. 7. Sätt enligt patentkravet 6, k ä n n e t e c k n a t av att hemocytlysatet härrör från en sötvattenskräfta, särskilt flodkräfta.7. A method according to claim 6, characterized in that the hemocyte lysate is derived from a freshwater crayfish, in particular a river crayfish. 8. Reagens för bestämning av bakteriellt endotoxin, k ä n n e t e c k n a t av att det innefattar (A) ett buffrat profenoloxidashaltigt blodkroppslysat från ett djur tillhörande någon av artropodklasserna kräftdjur (Crustacea) och insekter (Insecta), och (B) en detektorsubstans i form av åtminstone en fenolförening med förmåga att oxideras av enzymet fenoloxidas till en förening som kan påvisas fysikaliskt eller kemiskt, t.ex. spektro- fotometriskt eller kolorimetriskt.Reagent for the determination of bacterial endotoxin, characterized in that it comprises (A) a buffered propenol oxidase-containing blood cell lysate from an animal belonging to one of the arthropod classes crustaceans (Crustacea) and insects (Insecta), and (B) a detector substance in the form of at least a phenolic compound capable of being oxidized by the enzyme phenolic oxidase to a compound which can be physically or chemically detected, e.g. spectrophotometric or colorimetric.
SE8107599A 1981-12-17 1981-12-17 SETTING FOR DETERMINATION OF BACTERIAL ENDOTOXIN MEDIUM PROPHENOLOXIDAS-CONTAINING ANIMAL ANIMAL OR INSECTS AND REAGENTS SE431228B (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
SE8107599A SE431228B (en) 1981-12-17 1981-12-17 SETTING FOR DETERMINATION OF BACTERIAL ENDOTOXIN MEDIUM PROPHENOLOXIDAS-CONTAINING ANIMAL ANIMAL OR INSECTS AND REAGENTS
JP83500092A JPS58502082A (en) 1981-12-17 1982-12-17 Methods and reagents for the detection of bacteria and fungi
EP83900080A EP0096689A1 (en) 1981-12-17 1982-12-17 Method and reagent for detection of endotoxines or beta-1,3 glucanes from fungus or bacteria
PCT/SE1982/000430 WO1983002123A1 (en) 1981-12-17 1982-12-17 METHOD AND REAGENT FOR DETECTION OF ENDOTOXINES OR 'beta'-1,3 GLUCANES FROM FUNGUS OR BACTERIA

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SE8107599A SE431228B (en) 1981-12-17 1981-12-17 SETTING FOR DETERMINATION OF BACTERIAL ENDOTOXIN MEDIUM PROPHENOLOXIDAS-CONTAINING ANIMAL ANIMAL OR INSECTS AND REAGENTS

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SE8107599L SE8107599L (en) 1983-06-18
SE431228B true SE431228B (en) 1984-01-23

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