Evaluation of a Simple Method for Determination of IgG Titre
Anti-A or -B in Cases
of Possible ABO Blood Group Incompatibility

Reesink, H.W.; Hart, Mia van der; Loghem, J.J. van

doi:10.1159/000464537

Cited by 6 publications

(4 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Equal volumes of 2ME and serum were incubated at 370C for two hours. The treated sera were used without dialysis according to the method of Reesink et al (1972).…”

mentioning

confidence: 99%

Measuring IgG anti-A/B titres using dithiothreitol (DTT).

Knight¹

1978

J Clin Pathol

View full text Add to dashboard Cite

SUMMARY In comparing the use of the sulphydryl compounds 2-mercaptoethanol (2ME) and dithiothreitol (DTT) a rapid one-stage technique for titrating IgG anti-A and anti-B using DTT in an antiglobulin test was tried. The results correlated well with those obtained by either of the two-stage methods using 2ME or DTT. Bowley (1969) showed that the anti-A/B titre determined by a spin antiglobulin technique after 2ME inactivation correlated well with clinical severity in cases of ABO HDN, a titre of more than 1 in 256 being highly significant. 2ME has the following disadvantages that preclude its widespread use in routine laboratories: (1) the technique is time consuming, especially if dialysis is used; (2) without dialysis false-positive results may occur when using an anti-human globulin (AHG) technique; (3) 2ME has a pungent odour.DTT does not have these disadvantages since it is odourless, dialysis is not required before AHG techniques, and it is less time consuming. The period of inactivation can be reduced to as short as 15 minutes (Pirofsky and Rosner, 1974). The use of DTT in determining IgG anti-A/B titres is considered in this paper.Received for publication 27 July 1977 Methods 2ME was diluted to 0 1 M in phosphate buffered saline pH 7-4. Equal volumes of 2ME and serum were incubated at 370C for two hours. The treated sera were used without dialysis according to the method of Reesink et al. (1972).DTT We use a Technicon BG-10 grouping machine for routine ABO and Rh grouping, so in order to select sera for this study samples were also tested with A1B cells at 370C and those showing haemolysis within 30 minutes used. As all sera were tested within 24 hours of collection and only occasionally stored overnight at 4VC no additional source of complement was used. Other sera used were those containing antibodies that had been detected in the laboratory during routine testing and stored at -20'C until required.The broad spectrum AHG was obtained from Biotest Folex Limited and the anti-complement and anti-IgG from the Blood Group Reference Laboratory, London. All were used by a tile technique.

show abstract

“…Equal volumes of 2ME and serum were incubated at 370C for two hours. The treated sera were used without dialysis according to the method of Reesink et al (1972).…”

mentioning

confidence: 99%

Measuring IgG anti-A/B titres using dithiothreitol (DTT).

Knight¹

1978

J Clin Pathol

View full text Add to dashboard Cite

show abstract

“…Results were read with the help of a lamp provided with a magnifying mirror. Levels of IgG anti-A and anti-B in the sera of the mothers were determined by the indirect antiglobulin test using sera treated with 2-mercapto-ethanol [6]. In some cases, IgG anti-A/B levels were serologically determined directly on IgG fractions isolated from sera by ion-exchange chromatography using DEAE-cellulose (DE-52, Whatman Biochemicals).…”

Section: Serological Testsmentioning

confidence: 99%

Plasma Fibrinogen Concentration in ABO-Hemolytic Disease of the Newborn

Romano

Linares

Suárez

1982

Int Arch Allergy Immunol

View full text Add to dashboard Cite

Plasma fibrinogen and total serum proteins were measured in infants affected by ABO-hemolytic disease and in normal infants’ cord blood and blood taken 48 h after birth. The average plasma fibrinogen concentrations in the blood of infants with ABO-hemolytic disease was 447.36 mg/dl as compared with 267.09 mg/dl (p < 0.005) in cord blood and 313.82 mg/dl (p < 0.005) in blood samples taken 48 h afterbirth. A simple ‘autoagglutination’ test is found to be helpful when establishing the diagnosis of ABO-hemolytic disease.

show abstract

“…In order to determine the actual concentration of the clinically significant IgG antibodies, there is need to inactivate the IgM antibodies. Literature provides methods to inactivate IgM antibodies such as heat inactivation (HI) and use of sulfhydryl reagents such as 2‐mercaptoethanol (ME) and dithiothreitol (DTT; also called Cleland’s reagent) [9,10]. IgG antibodies are less susceptible to these treatments because the disulphide bonds between chains are not as labile as those of IgM subunits [11], although it is known that they can be slightly affected [12].…”

Section: Introductionmentioning

confidence: 99%

A prospective, observational study to compare effect of DTT treatment and heat inactivation on ABO isoagglutinin titers in O blood group donors

Pandey

Setya

Ranjan

et al. 2021

ISBT Science Series

View full text Add to dashboard Cite

Background and aims In order to determine actual concentration of clinically significant IgG antibodies, there is a need to inactivate IgM antibodies, which can be performed by DTT or heat. The aim of the present study was to compare the effect of DTT treatment and heat inactivation (HI) on ABO isoagglutinin titres performed by column agglutination technology (CAT) and conventional tube technique (CTT). Materials and methods This was a prospective, observational study conducted from October 2018 to March 2020. All consecutive O group donors were included. Serum from each donor was treated with DTT, and heat and tests were performed by CTT and CAT before and after treatment. Results A total of 2005 donors were included. 401 (20%) samples had inter‐observer variation by CTT, and 69 (3·4%) had inter‐observer variation by CAT. Median anti‐A and anti‐B titre results were higher with CAT as compared to CTT. Median anti‐A and anti‐B IgG titre results showed a one‐ to twofold difference between CTT and CAT with higher titres reported by CAT. Median IgM titres decreased two‐ to threefold after DTT treatment, whereas onefold decrease was confirmed after HI. Concordance was found to be poor. However, correlation was found to be strong. Conclusion There is overestimation of ABO IgG antibody titres and hence, estimation of ABO IgG titres after inactivation of IgM antibodies is strongly recommended. DTT treatment was found to be superior to HI in terms of efficiency in elimination of IgM activity.

show abstract

Evaluation of a Simple Method for Determination of IgG Titre Anti-A or -B in Cases of Possible ABO Blood Group Incompatibility

Cited by 6 publications

References 12 publications

Measuring IgG anti-A/B titres using dithiothreitol (DTT).

Measuring IgG anti-A/B titres using dithiothreitol (DTT).

Plasma Fibrinogen Concentration in ABO-Hemolytic Disease of the Newborn

A prospective, observational study to compare effect of DTT treatment and heat inactivation on ABO isoagglutinin titers in O blood group donors

Contact Info

Product

Resources

About