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At least 19 recordsLinked to original sources
PubMed2005

Serum protein media are important factors in the manual hexadimethrine bromide (polybrene) test, experience in China.

BACKGROUND AND OBJECTIVES: The use of the manual hexadimethrine bromide (polybrene) test in routine cross-matching after accurately detecting cell grouping and irregular antibodies is prevalent in China. This article reports the importance of serum protein mediums in the performance of the manual hexadimethrine bromide test. MATERIALS AND METHODS: Blood group O red blood cells and Blood group AB and Rh positive serum were collected at random from healthy blood donators, IgG anti-D serum separated from pregnant woman, then tested with each other by the manual hexadimethrine bromide methods in routine tests and some designed corresponding tests with IgG, IgM anti-D monoclonal diagnostic reagents and some serum protein components. RESULTS: Red blood cells that were adjusted to 3-5% suspension by normal saline then only added in 0.7 ml low ionic medium (LIM) and two drops of polybrene solution adhere to the surface of test tubes' bottom when centrifuged, so it was difficult to perform the next approach, but the adherence disappeared when red blood cells' concentrations exceeded 20-30%. Rh positive red blood cells coated by anti-D have the same phenomenon. This adherence can be prevented by serum medium diluted from 1:128 to 1:1024 times by normal saline and hemoglobin medium diluted from 1:32 to 1:128 times, but not by albumin or immunoglobulin medium. The denary logarithm values of the greatest inhibited dilutions of serum and hemoglobin elution between antibody sensitizing red blood cells and the same pre-sensitizing red blood cells tests were no significant difference (P value > 0.05). CONCLUSIONS: The whole serum or serum protein mediums are important factors that can influence successfully performance of the manual hexadimethrine bromide test. So appliance of the manual hexadimethrine bromide test to immunohematology laboratory, such as when performing titrations of serum or plasma, or when testing eluates for antibody activity, this adherence must be considered.

Artifacts↗
PubMed1987

Use of hexadimethrine bromide as a heparin-neutralizing agent in canine plasma.

Hexadimethrine bromide was evaluated as a heparin-neutralizing agent in a simple modification of the activated partial thromboplastin time test in canine plasma. Addition of various amounts of heparin in vitro to canine plasma indicated that heparin could be neutralized by adding 0.5 micrograms of hexadimethrine bromide 15 s before CaCl2 was added to the reaction mixture of the activated partial thromboplastin time test. In 8 dogs given (subcutaneous injection) 500 USP units of sodium heparin/kg, marked individual variations in clotting time prolongations were observed over the 12-hour period of study. The hexadimethrine bromide modification effectively neutralized the heparin-related clotting time prolongations to values that were not significantly different from base-line (preheparin) activated partial thromboplastin time values. The modification seems to be useful in confirming the presence of heparin and in monitoring heparin therapy in dogs.

Animals↗
PubMedDate not supplied

Comparison of a modified manual hexadimethrine bromide (Polybrene) and a low-ionic-strength solution antibody detection technique.

Manual hexadimethrine bromide (Polybrene) tests (Polybrene in low-ionic medium) were used in parallel with manual low-ionic-strength solution (LISS) procedures for the routine testing of patient samples referred to a general hospital blood bank. Of 5646 consecutive sera tested, 5167 (91.5%) did not react with either technique; 320 sera (5.7%) reacted in both methods. The Polybrene technique detected 63 antibodies which did not react in the LISS methods. One hundred sera did not react in the Polybrene test, but did react in the LISS methods. Sera showing discrepant results between the two methods were further tested in a reference laboratory. Polybrene tests appeared to be better in avoiding reactions due to clinically nonsignificant antibodies. The LISS methods, however, appeared to be more sensitive in detecting antibodies of potential clinical significance.

Blood Banks↗
PubMedDate not supplied

Evaluation of the manual hexadimethrine bromide (Polybrene) technique in the investigation of autoimmune hemolytic anemia.

The use of the direct manual hexadimethrine bromide (Polybrene) test (DPT) in the investigation of patients for autoimmune hemolytic anemia (AIHA) was evaluated. Seventy-nine blood samples from 68 patients were tested. A direct antiglobulin test (DAT) using monospecific reagents and the DPT were performed, and a concentrated ether eluate was tested. The DAT was positive in 62 (78%) of 79 patients and negative in 17 (22%). There is a good correlation among DAT, eluate, and DPT in demonstrating the presence of immunoglobulin on the red cell surface. In contrast, the DPT does not detect C3d and is often negative in cases of AIHA in which C3d alone is demonstrated by the DAT. In DAT-negative cases, DPT results correlated with reactive eluates. However, in four cases of steroid-responsive, DAT-negative hemolytic anemia, the DPT supported the diagnosis of AIHA when the eluate did not react. The DPT is a useful additional screening test for the investigation of AIHA, but it is not recommended as a replacement for either eluate testing or the DAT.

Anemia, Hemolytic, Autoimmune↗
PubMedDate not supplied

Red cell phenotyping using hexadimethrine bromide (Polybrene) in a microplate system.

To provide rapidly phenotyped units of blood, we adapted the hexadimethrine bromide (Polybrene) technique to microplate technology. Pilot samples from 282 donor units were phenotyped for antigens in the Rh, Kidd, Kell, Duffy, and Ss systems with a standard tube-testing method and a Polybrene-microplate (P-MP) technique. Diluted antisera and a 1 percent red cell suspension were used to give P-MP reactions that were accurate and easy to interpret. One microplate, containing 96 tests, was prepared and read within 15 minutes for P-MP tests yielding direct agglutination (Rh, Jkb, Fya, Fyb), or within 19 minutes for P-MP tests requiring an antiglobulin phase (Jka, K, S, s). No false-positive results were found. No false-negative reactions were found in typing for Rh, K, S, s, or Jka antigens. Phenotyping for Jkb and Duffy antigens gave a false-negative rate of less than 0.015. Considerable savings in reagents were obtained through microplate miniaturization and through the enhancement of apparent antibody avidity in the Polybrene/low ionic medium which permitted dilution of reagent antisera. The P-MP technique affords a rapid, accurate, simple, and inexpensive means of phenotyping large numbers of donor units.

Blood Group Antigens↗
PubMed1970

Effect of anti-proteases and hexadimethrine bromide on the release of a bradykinin-like substance during heating (46 degrees C) of rat paws.

1. The conditions in which the release of an active, bradykinin-like agent occurred when rat paws were heated to 46 degrees C were studied by means of the double coaxial perfusion of the subcutaneous spaces.2. The active material thus released stimulated the isolated rat uterus, produced a relaxing effect on the isolated rat duodenum, was destroyed by incubation with chymotrypsin and was potentiated by bradykinin-potentiating factor. LSD-25, in doses sufficient completely to block 5-hydroxytryptamine, did not affect the responses of the isolated uterus to the active material.3. The effects on this release of anti-proteases and hexadimethrine bromide, atropine and diphenhydramine were studied.4. Soy-bean trypsin inhibitor and hexadimethrine bromide added to the perfusion fluid produced a potent and reversible inhibition of the release of the active material; aprotinin and Kunitz inhibitor caused a temporary block.5. When administered intravenously, much larger doses of the substances were necessary to produce a similar block.6. Pretreatment of the animals with atropine plus diphenhydramine did not affect the release of the active kinin(s).7. Ligature of one iliac artery was followed by disappearance of the active material in the perfusate from the corresponding paw.8. These facts suggest that heating elicits a process leading to plasma extravasation and that the subcutaneous tissue is the chief site of release of the active material.

Amines↗
PubMed1964

EFFECT OF HEXADIMETHRINE BROMIDE ON PLASMA KININ FORMATION, HYDROLYSIS OF P-TOSYL-L-ARGININE METHYL ESTER AND FIBRINOLYSIS.

The antiheparin agent hexadimethrine bromide, in concentrations of 20 to 200 mug/ml., inhibited the activation by active Hageman factor of the plasma enzyme which releases kinin from substrate. Once activated, this kinin-forming enzyme was not consistently inhibited by hexadimethrine in a concentration of 1 mg/ml. Surfaces which induce kinin formation by activating Hageman factor in plasma (glass, kaolin, celite, barium carbonate and carboxymethylcellulose) were inactivated by bathing in aqueous solutions of hexadimethrine. The effects of hexadimethrine on Hageman factor and on glass were not abolished by amounts of heparin which neutralize most other actions of hexadimethrine. Hexadimethrine prevented the activation by kaolin, but not by streptokinase, of p-tosyl-L-arginine methyl ester-splitting and fibrinolytic factors in plasma; once these enzymes were activated by kaolin, they could not be inhibited by hexadimethrine. Hexadimethrine, given locally or intravenously into guinea-pigs, reduced the increase in capillary permeability produced by intracutaneous injections of kaolin suspensions.

Arginine↗
PubMedDate not supplied

The manual hexadimethrine bromide (Polybrene) test. Effects of serum proteins and practical applications.

Serum proteins affect the performance of the manual hexadimethrine bromide (Polybrene) test. Red cells suspended in the low-ionic medium adhere to the surface of test tubes when centrifuged. This adherence is prevented by serum components apoprotein B, fibronectin, von Willebrand factor, or serum diluted less than 75-fold, but not by serum albumin or immunoglobulin fractions. A high concentration of serum reduces the sensitivity of the test and destabilizes antibody-dependent aggregation. Disaggregation time (DT), defined as the time required for antibody-dependent aggregates to disperse, was used as an indicator of agglutination stability. DT was often shortened when antibodies were in native serums, but could be prolonged if serums were diluted with 0.85 percent saline. Other factors that influenced DT included antibody concentration, temperature, specificity, and sources of antibodies within specificities. When antibodies were present in a mixture, they produced various DTs related to individual specificities. Determination of these differences allowed the successful identification of multiple antibodies. The procedure, called differential disaggregation time, was also used for typing red cells coated by autoantibodies. Additional modifications improved the performance of the antiglobulin phase.

Antibody Specificity↗
PubMedDate not supplied

Anti-Jka, -C, and -E in a single patient, initially demonstrable only by the manual hexadimethrine bromide (Polybrene) test, with incompatibilities confirmed by 51Cr-labeled red cell studies.

Published reports have confirmed the superior sensitivity of the manual hexadimethrine bromide (Polybrene) test (MPT) for demonstrating many alloantibodies in vitro; however, the clinical significance of alloantibodies demonstrable exclusively by MPT has not been shown conclusively. A patient with macroglobulinemia experienced chills, fever, hemoglobinemia, and hemoglobinuria following the transfusion of 1 unit of red cells (RBCs) shown to be compatible by the low-ionic-strength antiglobulin (LIS-AG) method. Serologic investigation was negative. Intravascular hemolysis occurred with a second "compatible" unit. Serologic studies were again negative by LIS-AG and ficin-AG methods, but revealed anti-Jka by MPT. Both donors were Jk(a+b-), and 51Cr studies of the second donor's RBCs revealed a t1/2 of less than 30 minutes, with marked intravascular hemolysis. A LIS-AG-compatible Jk(a-) unit was transfused uneventfully, but with no rise in hematocrit. MPT next revealed anti-C; subsequent 51Cr studies with the Jk(a-), Cc donor's RBCs showed a 51Cr t1/2 of 100 minutes with slight intravascular lysis. Four transfusions of Jk(a-), C- blood were uneventful, but 5 days later the patient's hemoglobin declined. The following day, anti-E was demonstrable exclusively by MPT. 51Cr-labeled Jk(a-), C-, E- RBCs had normal 24-hour survival. The patient's hemoglobin rose to 11 g per dl following transfusions of Jk(a-), C-, E- RBCs, and he was discharged. In vitro studies employing the patient's purified IgM paraprotein revealed no interference with alloantibody binding or detection.

Aged↗
PubMedDate not supplied

Comparison of a commercial hexadimethrine bromide method and low-ionic-strength solution for antibody detection with special reference to anti-K.

The sensitivities of manual low-ionic hexadimethrine bromide (Polybrene, LIP) and low-ionic Polybrene indirect antiglobulin tests (LIPAT) were compared with those of a manual low-ionic-strength indirect antiglobulin test (LISS) by using a commercial Polybrene kit. One hundred antibodies were coded, titrated, and tested in parallel. LIP did not detect 36 antibodies: 31 anti-K, two anti-E, two anti-Fya, and one anti-Jka. LIPAT did not detect seven anti-K, two anti-E, and two anti-Jka. The combination of LIP and LIPAT did not detect two anti-E that were reactive only in a two-stage enzyme test and seven anti-K that had titers of 2 or lower by LISS. LISS detected all antibodies except for the two enzyme-reactive anti-E. There were no significant differences in the titers of 63 percent of the antibodies studied. For 54 percent of the antibodies in the Kell system, LISS produced significantly higher titers; for 25 percent of antibodies in the Rh system, LIP did so. The poor sensitivity of the Polybrene kit for anti-K makes it unsuitable as a primary method for antibody screening.

Coombs Test↗
PubMedDate not supplied

A proposal for compatibility testing incorporating the manual hexadimethrine bromide (Polybrene) test.

In November 1984, the Standards Committee of the American Association of Blood Banks changed the requirements for pretransfusion testing by making the performance of an antiglobulin crossmatch optional when the antibody screening test is negative. The crossmatch would be necessary only to confirm ABO compatibility. Many will welcome this change; others will persist in their current methods. This article presents data supporting the use of the manual hexadimethrine bromide (Polybrene) test, a 1-minute room temperature procedure, as a crossmatch technique when the antibody screening test is negative. The manual Polybrene test (MPT) is an effective method for detecting ABO incompatibility. Forty-seven randomly selected serums gave expected results with A1, A2, and B red cells. Only 66 percent of 84 group B sera were serologically incompatible with A2B red cells by MPT, but the same results (69% positive) were observed using a 5-minute low-ionic-strength solution (LISS) room temperature technique. As only 37 percent of these crossmatches were incompatible using a LISS immediate spin (IS) method, the reliability of an IS method is questioned. An MPT crossmatch provides added security in that most unexpected blood group antibodies are demonstrable by this method. Of 106 serums tested which contained antibodies, 83 reacted. We believe that the MPT provides a rapid and sensitive test that, accompanied by a carefully performed antibody screening test, meets the requirements of Standards and will provide for safe red cell transfusion without the need for an antiglobulin crossmatch.

ABO Blood-Group System↗