Flocculation Tests and their Application to the Study of Liver Disease

Reinhold, John G.

doi:10.1016/s0065-2423(08)60099-2

Cited by 10 publications

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“…Chronic infection, for instance syphilis, is one of the conditions associated with increased thymol turbidity (TT) of serum (Weiss, 1969; Menke, 1975).Factors contributing to the TT are discussed by Reinhold (1960). The free lipids of serurm play a part, but proteins have also been found to react with the thymol reagent (Kunkel and Hoagland, 1947;Franklin, 1959).…”

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Role of IgG fractions with high isoelectric points in the thymol turbidity test in syphilis. Evidence for an increase in basic IgG in early syphilis.

Sluis¹,

Menke²

1975

Sexually Transmitted Infections

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The thymol turbidity test (TTT) is a non-specific reaction, which gives results above normal in a variety of diseases (Stillerman, 1948). Chronic infection, for instance syphilis, is one of the conditions associated with increased thymol turbidity (TT) of serum (Weiss, 1969; Menke, 1975).Factors contributing to the TT are discussed by Reinhold (1960). The free lipids of serurm play a part, but proteins have also been found to react with the thymol reagent (Kunkel and Hoagland, 1947;Franklin, 1959).Electrophoresis shows that the proteins precipitated by the thymol reagent behave like the beta and gamma proteins. The addition of gamma globulins to normal serum is followed by an increase in TT, and the removal of these proteins from positive TTT sera is followed by a negative reaction (Kunkel and Hoagland, 1947).As Franklin (1959) has shown, basic fractions of the gamma region are important in the reaction. Although negative when tested alone, these fractions affect the TT when tested in the presence of lipoproteins. Hartmatn, Viallet, and Fauvert (1963) described the so-called 'starter mechanism' of IgM. The addition of IgM myeloma proteins to normal serum was followed by an increase in TT, an effect which was not observed with IgG. However, when both IgM and IgG were added, the increase was larger than could be expected from the amount of IgM alone. These results indicate that some of the immunoglobulins play a part in the TTT. Since we were interested in the course of immunological events taking place after infection with T. pallidum, we investigated some of the factors contributing to the increased thymol turbidity in syphilis. In the experiments described in this communication, we studied the action of IgG isolated from syphilitic sera in the TTT. Material aind methodsAll the chemicals used were of analytical reagent grade. Antisera were obtained from the Central Laboratory of the Dutch Red Cross (Amsterdam, The Netherlands) and Received for publication August 30, 1974 Nordic (Tilburg, The Netherlands). Sera from syphilitic subjects were used either at once or after storage at -700C. for varying lengths of time. In all cases the diagnosis was confirmed by a positive darlfield examination and/or a positive Treponema pallidum immobilization (TPI) reaction. The TTT was performed by diluting 0-1 ml. of serum with 6 ml. of thymol reagent pH = 7-55 to 7 60 (Baker, Deventer, The Netherlands).After 30 min. the turbidity was read on a Vitatron spectrophotometer at 637 nm. Protein content was measured by the biuret reaction, using Lab-Trol (Dade, Miami, Florida, USA) as a standard.Zone electrophoresis in Pevikon blocks was performed according to the method of Bocci (1964) in barbitalbarbital sodium buffer, pH = 8-6, ,u 0-05, in blocks measuring 36 x 16 x 1 cm. 7 to 8 ml. serum from a syphilitic patient was dialysed in barbiturate buffer and applied to the block in a groove 12 cm. from the cathodal end. A voltage gradient of 3 3 V/cm. was applied to the block.After passage of current for 28 hrs, the block was divide...

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“…Factors contributing to the TT are discussed by Reinhold (1960). The free lipids of serurm play a part, but proteins have also been found to react with the thymol reagent (Kunkel and Hoagland, 1947;Franklin, 1959).…”

mentioning

confidence: 99%

Role of IgG fractions with high isoelectric points in the thymol turbidity test in syphilis. Evidence for an increase in basic IgG in early syphilis.

Sluis¹,

Menke²

1975

Sexually Transmitted Infections

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“…Assay of patient sera and purified MPs for ZTT and TTT . The ZTT and TTT of patient serum or purified MP in phosphate-buffered saline at pH 7.0 (PBS) were carried out in a Hitachi 736-60 automatic analyzer by Reinhold's method (7) and the Shank-Hoagland method (8), respectively, using reagents purchased from Nissui Co. MP was isolated from patient serum dialyzed against Veronal buffer (pH 8.6) by zone electrophoresis (ZE) on agar plate and was purified by several repetitions of ZE, followed by gel-filtration on Sephacryl S-300.…”

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confidence: 99%

Studies on the Relationship between Serum Colloidal Reactions (ZTT and TTT) and IgG Subclasses, Especially IgG₁ and IgG₂

Miyagawa

Okamoto

Morita

et al. 1991

Microbiology and Immunology

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The relationship between zinc sulfate turbidity test (ZTT) and thymol turbidity test (TTT), and IgG subclasses, especially IgG1 and IgG2, was studied. Serum colloidal reactions of specimens from patients with IgG‐myeloma usually show abnormally high values in ZTT and low values in TTT. But in some cases. values in ZTT and TTT are both low, and in a few cases both values are abnormally high. In order to see the reason why IgG‐myelomas are classified into these three groups according to serum colloidal reactions, IgG subclasses of monoclonal proteins (MPs) isolated by zone electrophoresis were determined by immunodiffusion with anti IgG‐subclasses. Results revealed that IgG1‐myeloma showed high ZTT and low TTT and IgG2‐myeloma showed low ZTT and TTT. Only two out of our 29 cases with IgG‐myeloma showed high ZTT and TTT. MP from one of them belonged to IgG1‐k and that from the other to IgG2‐λ. We checked the relationship between turbidity tests and electrophoretic mobility of MP and also that between these tests and L‐chains' type of MP, but did not find any relationship between them. These findings suggest that IgG1 innately reacts with ZTT but not with TTT, while IgG2 does not react with any of these tests. MP from cases showing high values in both ZTT and TTT may have a special idiotype.

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“…Flocculation tests, such as the Takata-Ara, cephalin-colesterol, zinc sulphate and thymol turbidity have been used to replace the relatively arbitrary electrophoretic analysis of serum proteins (Maclagan, 1948;Reinhold, 1960). However, the flocculation tests developed for human serum have generally been less suitable for animal sera (Cornelius, 1957).…”

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confidence: 99%

Determination of γ‐globulin in dog serum by glutaraldehyde

Sandholm

Kivistou

1975

J of Small Animal Practice

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The serum y‐globulin level is a function of the production of immunoglobulins (activity of the lymphoid system) and their catabolism (catab‐olized by the liver). The demonstration of elevated serum y‐globulin is therefore helpful in detection of chronic infections and hepatic or biliary disorders. A semiquantitative test for y‐globulin in canine serum is described. Dilute glutaraldehyde solutions polymerize the most basic serum proteins (γ‐globulins) coagulating the serum. ‘The γ‐globulin test is performed simply by mixing the reagent’ (6 % glutaraldehyde) with serum in the proportion of 1:10. The coagulation time is inversely correlated to the γ‐globulin content in the sera. Hyper γ‐globulinaemic sera will coagulate within 3 minutes.

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Flocculation Tests and their Application to the Study of Liver Disease

Cited by 10 publications

References 204 publications

Role of IgG fractions with high isoelectric points in the thymol turbidity test in syphilis. Evidence for an increase in basic IgG in early syphilis.

Role of IgG fractions with high isoelectric points in the thymol turbidity test in syphilis. Evidence for an increase in basic IgG in early syphilis.

Studies on the Relationship between Serum Colloidal Reactions (ZTT and TTT) and IgG Subclasses, Especially IgG₁ and IgG₂

Determination of γ‐globulin in dog serum by glutaraldehyde

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Flocculation Tests and their Application to the Study of Liver Disease

Cited by 10 publications

References 204 publications

Role of IgG fractions with high isoelectric points in the thymol turbidity test in syphilis. Evidence for an increase in basic IgG in early syphilis.

Role of IgG fractions with high isoelectric points in the thymol turbidity test in syphilis. Evidence for an increase in basic IgG in early syphilis.

Studies on the Relationship between Serum Colloidal Reactions (ZTT and TTT) and IgG Subclasses, Especially IgG1 and IgG2

Determination of γ‐globulin in dog serum by glutaraldehyde

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Studies on the Relationship between Serum Colloidal Reactions (ZTT and TTT) and IgG Subclasses, Especially IgG₁ and IgG₂