A prospective study to determine the safety of omitting the antiglobulin crossmatch from pretransfusion testing

Transfusion medicine laboratories routinely perform a series of pretransfusion serological tests including: ABO grouping, Rh typing, and investigation of the recipient's serum to detect antibodies against blood group antigens (antibody screen). As a final check, most laboratories also perform a crossmatch in which the recipient's serum is incubated with the donor's red cells followed by the addition of an antiglobulin reagent (antiglobulin crossmatch). The need for the antiglobulin crossmatch when the antibody screen is negative has been questioned because there are few antibodies that are detected by this test. Such antibodies are usually directed against low incidence antigens that are not expressed on the screening cells and in many cases the clinical importance of these antibodies is uncertain. For these reasons, we performed a prospective study in which patients requiring red cell transfusion had a group and screen performed. If the antibody screen was negative the antiglobulin crossmatch was omitted. Following the transfusion of the blood, the antiglobulin crossmatch was performed to look for any potential incompatibility. All patients were monitored both serologically and clinically. Over the 2-year interval of the study 9128 patients were entered. There were 8936 patients (97.9%) with a negative antibody screen and 26.9% (2404 patients) were transfused a total of 10,899 red cell concentrates. The antiglobulin crossmatch performed after the transfusion indicated that 168 red cell concentrates (1.5%) would have been incompatible if the antiglobulin crossmatch had been performed pretransfusion. These 168 red cell concentrates were transfused to 119 patients during 130 transfusion episodes (defined as all transfusions administered within 24 h). Of the 130 transfusion episodes, 79.2% (103/130) were false positive laboratory results. There were 27 transfusion episodes where the antiglobulin crossmatch on blood transfused was positive due to an IgG antibody. Even though these transfused red cell concentrates were designated incompatible by the antiglobulin crossmatch, none of the patients receiving this blood had clinical or serological evidence of haemolysis. We concluded that the antiglobulin phase of the crossmatch can be omitted from pretransfusion testing without putting patients at risk.     

The UK National External Quality Assessment Scheme in Blood Group Serology. Compatibility testing 1983–1984: the influence of variables in test procedures on detection of incomplete antibodies

Summary Error rates in exercises in compatibility testing varied between 3% and 36% over the six years 1979–1984. Evolution of serological procedures was continuous through this period but without clear evidence of improvement in performance of antibody detection although performance in the UK appears to be comparable with that elsewhere. Relationships have been established between a number of variables of reagents and techniques, and their performance. The influence of some variables is reasonably clear but there appear to be interactions between certain variables in their effects on performance, and interpretation is less straightforward. A test for agglutination of enzyme treated cells together with an antiglobulin test appears to be the most reliable combination for detection of incomplete antibodies in compatibility testing although one stage enzyme techniques are an inevitable compromise between reliability and convenience. Compatibility testing should not be considered in isolation from antibody screening in determining the optimum combination of tests for pre-transfusion antibody detection.     

The UK National External Quality Assessment Scheme in Blood Group Serology. Compatibility testing 1983-1984: the influence of variables in test procedures on detection of incomplete antibodies

Error rates in exercises in compatibility testing varied between 3% and 36% over the six years 1979-1984. Evolution of serological procedures was continuous through this period but without clear evidence of improvement in performance of antibody detection although performance in the UK appears to be comparable with that elsewhere. Relationships have been established between a number of variables of reagents and techniques, and their performance. The influence of some variables is reasonably clear but there appear to be interactions between certain variables in their effects on performance, and interpretation is less straightforward. A test for agglutination of enzyme treated cells together with an antiglobulin test appears to be the most reliable combination for detection of incomplete antibodies in compatibility testing although one stage enzyme techniques are an inevitable compromise between reliability and convenience. Compatibility testing should not be considered in isolation from antibody screening in determining the optimum combination of tests for pretransfusion antibody detection.     

Standardizing Chlamydia pneumoniae assays: recommendations from the Centers for Disease Control and Prevention (USA) and the Laboratory Centre for Disease Control (Canada).

Chlamydia pneumoniae has been associated with atherosclerosis and several other chronic diseases, but reports from different laboratories are highly variable and "gold standards" are lacking, which has led to calls for more standardized approaches to diagnostic testing. Using leading researchers in the field, we reviewed the available approaches to serological testing, culture, DNA amplification, and tissue diagnostics to make specific recommendations. With regard to serological testing, only use of microimmunofluorescence is recommended, standardized definitions for "acute infection" and "past exposure" are proposed, and the use of single immunoglobulin (Ig) G titers for determining acute infection and IgA for determining chronic infection are discouraged. Confirmation of a positive culture result requires propagation of the isolate or confirmation by use of polymerase chain reaction (PCR). Four of 18 PCR assays described in published reports met the proposed validation criteria. More consistent use of control antibodies and tissues and improvement in skill at identifying staining artifacts are necessary to avoid false-positive results of immunohistochemical staining. These standards should be applied in future investigations and periodically modified as indicated.     

Serodiagnosis of tuberculosis: due to shift track

Development of novel diagnostics for tuberculosis has so far been governed by the clinical requirement of improving the detection of patients with paucibacillary forms of the disease. For this aim, serological assays have been evaluated using several antigens, but were found insufficiently sensitive, because antibody production associates with the bacterial load of the disease. Consequently, detection of antibodies against a relatively small number of selected well-defined antigens has a much higher sensitivity for sputum smear-positive pulmonary disease in adult HIV-negative patients. They are the most active in generating and spreading aerosols containing live tubercle bacilli, but their detection is often delayed, thus perpetuating the transmission of the infection and disease in the population. High volume throughput serological screening of clinical suspects with mild clinical symptoms may help to achieve diagnosis earlier, than currently used procedures. Such expanded testing could be done more efficiently in laboratories, than at 'points-of-care' and at a lower cost than other tests. The feasibility of this approach towards reducing the delayed diagnosis of the most infectious cases of pulmonary tuberculosis needs to be ascertained in prospective diagnostic trials, in populations at a high risk. Reducing the transmission of tuberculosis is of key importance for achieving its continued decline and therefore it is proposed, that the aims of serological screening should shift from clinical to public health priorities.     

Interlaboratory variation in the detection of HPA-specific alloantibodies and in molecular HPA typing

BACKGROUND AND OBJECTIVES: Platelet immunology quality assurance exercises have been organized by National Institute for Biological Standards and Control since 1991 and, as of 2006, 35 laboratories participate in the serology section. Molecular human platelet antigen (HPA) typing has been included in the exercises since 1998 and, as of 2006, 29 laboratories participate in this part of the exercise. This report details the performance of laboratories in these two areas. MATERIALS AND METHODS: Every 6 months laboratories are sent up to four coded serum/plasma samples for testing in their in-house assays for HPA antibody detection/identification and four coded whole blood samples to be typed for HPA-1, -2, -3 -5 and (since 2003) -15 by their molecular typing assays ('genotyping'). RESULTS: Fifty-two sera containing HPA-specific alloantibodies and 13 sera that were inert, contained only human leucocyte antigen (HLA) class I or high-titre anti-A+B antibodies were distributed; 15 sera were issued in more than one exercise. The percentage of participating laboratories that were able to detect HPA-specific alloantibodies ranged from 15.0 to 100%; the percentage that were able to correctly determine the specificities also ranged from 15.0 to 100%. Over the 20 serology exercises the percentage of laboratories classified as poor performers ranged from 3.1 to 36.7%. A total of 12 780 individual HPA genotyping results were assessed. The overall error rate was 0.7% but there was considerable variation between HPA alleles. Over 11 exercises the percentage of laboratories classified as poor performers varied from 6.3 to 26.3%. CONCLUSIONS: The ability to detect and to identify platelet-specific alloantibodies varied widely between laboratories and between various examples of antibodies issued. An increase in the number of laboratories screening for HPA-15 antibodies was seen, although detection and identification of these antibodies was problematic. The majority of examples of HPA-3a antibodies and some examples of HPA-1a and -5b were also difficult to detect and identify. In addition, this scheme has shown that despite the apparent reliability of molecular typing techniques, mistakes do occur, particularly with certain systems. Approximately one in five laboratories participating in the serology exercises and one in seven participating in the genotyping exercises were classified as poor performer at one point or more during the series of exercises.     

Pooled cells versus individual screening cells in pre-transfusion testing

Pooling of screening red cells used in pre-transfusion testing has been discouraged by many workers since it might lead to the failure to detect alloantibodies. However, there is minimal scientific data to support this view and in the UK a high proportion of blood transfusion laboratories were still using pooled red blood cells for antibody screening in 1982-1983. In order to solve the dilemma of sensitivity of serological tests when using pooled versus individual screening cells, a comparative evaluation of both approaches was carried out. One hundred and five sera known to contain weak, warm-reacting clinically significant alloantibodies were tested by the three same standard sensitive techniques using samples of individual screening cells and a pool of the same cells in parallel. Our results show that when pooled cells were used, 12% of the selected alloantibodies were undetectable and a further 23% were only detectable microscopically. All antibodies examined were easily detectable macroscopically when individual screening cells were used. It is concluded that the sensitivity of pre-transfusion screening tests is reduced when pooled red cells are employed as opposed to individual samples of screening cells and that the use of pooled cells should be discouraged in routine pre-transfusion and antenatal antibody screening.     

Report on the 14th international society of blood transfusion platelet immunology workshop

Background and Objectives The aims of the 14th ISBT Platelet Immunology Workshop were to evaluate in‐house methods for detection of antibodies to human platelet antigens, to compare the sensitivity and specificity of antibody detection using a panel of monoclonal antibodies and to evaluate genotyping methods and establish procedures for drug‐dependent antibody detection. Materials and Methods Forty‐two laboratories from 23 countries participated. Samples and reagents provided for the five different exercises. Results The ability of participating laboratories to correctly identify the HPA antibody specificity in the nine samples ranged from 20% to 97%. The greatest difficulty was observed with samples that contained antibodies against HPA‐3b and GPIV. The significant differences in optical density values by monoclonal antibody of immobilization of platelet antigens (MAIPA) assay were observed when testing the same platelet‐specific antibodies. HPA genotyping of DNA with novel mutations did not significantly affect the results. The overall average discrepancy rate was 2·15% for genotyping of 10 DNA samples from well‐characterized Epstein–Barr virus transformed cell lines. For detection of drug‐dependent antibodies, excellent results for specificity and sensitivity were obtained by the laboratories using the MAIPA and flow cytometry. Conclusions Most laboratories were able to identify the majority of HPA antibodies; however, significant disparities were observed in proficiency testing. MAIPA assay sensitivity is influenced by the monoclonal antibody clone used. DNA with new mutations and EBV cell lines are valuable samples to ensure accurate genotyping. A sensitive and specific drug‐dependent antibody assay performed well in the hands of participants.     

Quality control assessment of Canadian laboratories testing for Lyme disease

In June 1990 a quality control assessment was undertaken of Canadian public health laboratories testing for antibodies to Borrelia burgdorferi, the etiological agent of Lyme disease. Twenty sera were distributed to nine laboratories, including 12 obtained from patients in Lyme endemic areas and presumed to be serological positives, and eight prescreened negative controls. Seventeen serological reports were submitted, comprising nine enzyme-linked immunosorbent assays (elisa), six immunofluorescent assays and two Western blot assessments. Antibodies were detected in 11 of the 12 sera which had been presumed to be positive. Assuming 11 positive sera had been submitted, the test sensitivities varied from 88.9 to 100% by elisa, and 54.5 to 90.1% by immunofluorescent assay. Specificities were 100% for all but one elisa and one immunofluorescent assay assessment. The results indicate a satisfactory performance by elisa but a need for upgrading or replacement of some immunofluorescent assay tests.     

The mycobacteriology laboratory and new diagnostic techniques

Use of the most rapid and reliable laboratory tests for mycobacterial detection, identification, and susceptibility testing is important for TB control. In 1993, CDC experts made recommendations regarding optimal methods of mycobacterial testing (i.e., stains for AFB, culture, identification, and susceptibility testing of M. tuberculosis) and turnaround times for reporting results. Various technical advances have enhanced the diagnostic capability of the laboratory and/or improved laboratory efficiency since then. The commercial NAA tests for direct detection of MTBC have the greatest potential to impact patient care. To assist physicians, CDC experts have published recommendations concerning use of the NAA tests for management of patients with suspected TB, with emphasis on the MTD assay, which is approved for both AFB smear-positive and smear-negative specimens. With regard to mycobacterial culture, totally automated, nonradiometric systems are commercially available. For mycobacterial identification, various molecular techniques have been developed, but at present, they are used predominantly in research or large reference laboratories. Molecular tests also have proved useful for better understanding the epidemiology of TB and investigating episodes of suspected laboratory cross-contamination. With regard to mycobacterial susceptibility testing, use of the new automated culture systems for testing MTBC is under evaluation, but only one such system has been approved for this purpose. In addition, laboratory guidelines for susceptibility testing of MTBC and certain NTM have recently been published by the NCCLS.     

New technologies to replace current blood typing reagents

PURPOSE OF REVIEW: This review summarizes recent developments in blood grouping and compatibility testing in transfusion medicine. RECENT FINDINGS: Identification of the molecular characteristics of the major human blood groups has provided an opportunity to develop methods for blood group phenotyping using DNA-based technology. Various studies have demonstrated the feasibility of such an approach and have demonstrated the potential to change current procedures for identifying compatible blood, both in routine settings and in highly immunized patients, for whom compatible blood is difficult to obtain. In the obstetric setting, isolation of cell-free DNA from maternal plasma for fetal blood grouping provides a minimally invasive method for determining the risk for haemolytic disease in the newborn. Recombinant technology for synthesizing blood group proteins, although in its infancy, has the potential to change longstanding antibody identification procedures. SUMMARY: The molecular revolution occurring throughout medicine is broadly manifest in all areas of transfusion medicine and should contribute to transfusion safety.     

A comparison of two automated methods for the detection and identification of red blood cell alloantibodies

Background

The aim of this study was to compare the routine use of two automated systems (OrthoAutoVue Innova, microcolumn, and Immucor Galileo, solid phase) for the screening and identification of irregular red blood cell alloantibodies in samples, analysed in our Transfusion Service during 6 months of normal activity. The study focused particularly on an evaluation of the repeatability of the screening tests, the identification of antibody specificities and the identification of antibodies in samples showing discordant results.

Materials and methods

Overall 2,229 samples from potential blood donors (A), multiply transfused patients with blood disorders (DH), potential transfusion recipients (TS), and external cases (E) were studied. The protocols were carried out according to the manufacturers recommendations.

Results

The screening tests detected 78 samples that were positive with both systems, while 18 were positive only with Immucor and 11 only with Ortho (thus, overall, Immucor detected 96 positive samples and Ortho 89 positive samples). The use of the respective identification panels enabled us to identify the antibodies in 65 samples with Immucor and in 61 samples with the Ortho system; 74 antibodies were identified with Immucor (55 with a single specificity and 19 with mixed specificities) and 68 antibodies with Ortho (51 and 17, respectively). In the remaining cases (31 samples for Immucor and 28 for Ortho), the antibody specificity was not identified. The two systems were found to be essentially similar. The Immucor system revealed a greater number of antibodies, mainly because of its greater sensitivity at detecting anti-D antibodies.

Conclusions

Both systems showed a repeatability of over 85%, demonstrating that automation of immunohaematological tests is advantageous. The specificity of the antibody was identified in 68% of the samples. Furthermore, using the two systems led to the identification of ten new antibodies (6 anti-D, 2 anti-E, 1 anti Le^a, and 1 anti-Vel), which would not have been detected had only one of the two methods been used.     

Report on the 13th International Society of Blood Transfusion Platelet Immunology Workshop

BACKGROUND AND OBJECTIVES: The aim of the 13th International Society of Blood Transfusion Platelet Immunology Workshop was to compare the sensitivity and specificity of the in-house method for the detection of human platelet antigen (HPA) antibodies currently used in participating laboratories with a modified rapid protocol for the monoclonal antibody (mAb) immobilization of platelet antigen (MR-MAIPA) assay. MATERIALS AND METHODS: Twenty-eight laboratories from 15 countries participated. A set of four freeze-dried minimum potency reference reagents with known single-specificity HPA antibodies were supplied for testing by titration with both assays and two coded freeze-dried plasma samples were provided for antibody specificity testing. Critical reagents and materials for the MR-MAIPA were provided including lyophilized panel platelets and five capture mAbs. RESULTS: Titration of the reference standards showed that the sensitivity of the MR-MAIPA was the same as the in-house methods. The proposed replacement anti-HPA-1a reference reagent 05/106 gave results that did not differ significantly from the current reference reagent 93/710. The results with the two blinded samples showed that in the first sample, 27 out of the 28 laboratories were able to correctly identify the anti-HPA-1a present when using their respective in-house methods, but only 23 correctly identified the antibody when using the rapid MAIPA method. The results from the second sample, which contained multispecificities, showed that only 50% of the participants correctly identified all five antibodies present using their in-house method. The results for the rapid MAIPA were lower, with only 32% identifying all specificities. The variability in the reconstitution of the freeze-dried platelets may have been one of the contributing factors to the poorer results. CONCLUSIONS: The sensitivity of the MR-MAIPA compared favourably with that of the in-house methods. Most laboratories were able to identify anti-HPA-1a alone in Sample 1 but more than half of the participants were not able to correctly assign the specificity of all HPA antibodies present in the second sample. The usefulness of the panel of freeze-dried platelets varied considerably between laboratories.     

The UK National External Quality Assessment Scheme in Blood Group Serology. Compatibility testing 1981–1982: performance and practice

Summary. The design of exercises of compatibility testing was modified in 1981 in order better to accommodate participants' serological practices. Ten reference laboratories were also enrolled in order to determine the ‘correct’ results for each exercise. In 1981–1982 3.5 to 36% of participants missed incompatibilities in exercises in which undiluted antibodies were issued and this did not represent an improvement over performance obtained in 1979–1980. Surveys were undertaken of antiglobulin test procedures and revealed that serological practices continue to change, old techniques are being modified, new techniques are being employed but standardization shows little overall improvement. Surveys of quality control procedures and of cross-match procedures for agglutination in albumin and for agglutination of enzyme treated cells show equal lack of standardization.     

The UK National External Quality Assessment Scheme in Blood Group Serology. ABO and D grouping and antibody screening 1982-1983

In seven surveys of blood grouping the overall rates of major error were 0.12% and 0.37% for uncomplicated ABO and D grouping respectively. Of 17 errors of ABO grouping, 13 were errors of transposition or interpretation and four were apparently technical. Of 52 errors of D grouping, 20 appeared to be errors of transposition or interpretation and 32 were apparently technical. Of the 32 technical errors of D grouping, 31 were D-negative grouped as Du (29) or D-positive (2) and most of these errors were due to misgrouping in the antiglobulin test. Causes of error in D grouping by antiglobulin test include anti-Bg and other contaminating immune antibodies, residual unabsorbed anti-A and the inherently high rate of false positive results obtained in the antiglobulin test. In view of the lack of benefit of Du testing to blood recipients or to pregnant women and of the possible adverse consequences of misgrouping D-negative patients as Du or D-positive, it is recommended that Du testing be abandoned in these groups of patients. The surveys of antibody screening demonstrated lack of standardisation and error rates similar to those previously reported in the UK for compatibility testing.     

Detection of Irregular Red Cell Antibodies: More than 3 Years of Experience with a Gel Technique and Pooled Screening Cells

Background and objectives: The purpose of this study was to evaluate more than 3 years of experience with a gel technique in combination with pooled screening cells for the detection of irregular red cell antibodies. Materials and methods: Conventional serologic methods were used for blood typing, antibody screening and cross-matching until the end of 1992. We introduced the gel technique as a routine assay for antibody detection and identification in 1993. Results: After the tube technique had been abandoned, the number of false-positive antibody screening tests was reduced by 71%, positive antibody screening tests by 33%, enzyme agglutination by 100% and rouleaux reactions and cold-reacting antibodies by more than 50%. There was a 40% increase in first-time detection of clinically relevant antibodies. We saw no increase in delayed haemolytic transfusion reactions. Conclusions: For the detection of irregular red cell antibodies, pooled screening cells in combination with a gel technique are at least as efficient and safe as a conventional tube technique with unpooled test cells.     

Evaluation of serological diagnosis tests for tuberculosis in hemodialysis patients

Patients receiving hemodialysis are generally considered to be at increased risk of developing tuberculosis. In the current study, in order to evaluate the usefulness of serological tests in dialysis patients, serum antibodies for tuberculous glycolipids antigen (TBGL) and for lipoarabinomannan (LAM) were measured in hemodialysis patients. The present study included 243 hemodialysis patients. Serum antibodies for TBGL and LAM were measured. Tuberculin skin tests were carried out and chest X-rays evaluated at the same time. There were no patients with active tuberculosis at the time of blood sampling. Thirty-six patients (14.8%) and 25 patients (10.3%) were positive for anti-TBGL antibody and anti-LAM antibody, respectively. One hundred and fifty-five patients (63.8%) were positive for tuberculin skin testing and 123 patients (50.6%) had old pulmonary tuberculosis on their chest X-ray. There was no significant correlation between the results of anti-TBGL antibody and anti-LAM antibody. There were no relationships among the results of tuberculin skin test and the two serological tests. However, positivity of anti-TBGL antibody and anti-LAM antibody was significantly higher in patients with findings of old tuberculosis on the chest X-ray than those without findings. The current results show that these serological tests are positive more frequently in hemodialysis patients without any proof of active tuberculosis than in healthy subjects (2%) and careful interpretation is necessary for relevant results.     

Antiphospholipid syndrome: laboratory testing and diagnostic strategies

The antiphospholipid syndrome (APS) is diagnosed in patients with recurrent thromboembolic events and/or pregnancy loss in the presence of persistent laboratory evidence for antiphospholipid antibodies. Diagnostic tests for the detection of antiphospholipid antibodies include laboratory assays that detect anticardiolipin antibodies, lupus anticoagulants, and anti-β(2)-glycoprotein I antibodies. These assays have their origins beginning >60 years ago, with the identification of the biologic false positive test for syphilis, the observation of "circulating anticoagulants" in certain patients with systemic lupus erythematosus, the identification of cardiolipin as a key component in the serologic test for syphilis, and the recognition and characterization of a "cofactor" for antibody binding to phospholipids. Although these assays have been used clinically for many years, there are still problems with the accurate diagnosis of patients with this syndrome. For example, lupus anticoagulant testing can be difficult to interpret in patients receiving anticoagulant therapy, but most patients with a thromboembolic event will already be anticoagulated before the decision to perform the tests has been made. In addition to understanding limitations of the assays, clinicians also need to be aware of which patients should be tested and not obtain testing on patients unlikely to have APS. New tests and diagnostic strategies are in various stages of development and should help improve our ability to accurately diagnose this important clinical disorder.     

Prenatal serologic screening in Bahrain.

Between 1988 and 1990, serological surveys designed to study local disease prevalence and assess the clinical value of various prenatal screening tests were undertaken at Salmaniya Medical Center in Bahrain. High maternal antibody prevalence (greater than 85%) to cytomegalovirus (CMV), herpes simplex virus (HSV) and rubella was demonstrated, and 28% showed antibodies to Toxoplasma gondii. The lowest seroprevalence values were found for HBsAg (1.2%) and Treponema pallidum (0.9%). Routine testing for rubella, syphilis, and hepatitis B are advocated for all pregnancies in Bahrain. In contrast, CMV and HSV serologies are not recommended. Toxoplasma antibody testing remains controversial, but the lack of a proven agent to prevent congenital toxoplasmosis coupled with the high cost of serial testing mitigates against its routine use at this time.     

Automated Red Cell Antibody Analysis. A Parallel Study

Abstract. The AutoAnalyzer method of red blood cell antibody detection can be adapted to identify the serological specificity of these same antibodies. In the present study, 1,152 AA-positive sera obtained during AA screening were investigated. In 217 (19%) of these sera blood group antibodies were identified by manual techniques. Since the AA techniques are generally more sensitive than manual methods an attempt was made to identify antibodies by the AA methods in 823 sera which were negative manually. The low-ionic strength-Polybrene (LISP) method was found to be suitable for antibody identification because of its sensitivity in the detection of most red cell antibodies and the small technical deviations of OD from the base-line. The bromeline-methyl cellulose (BMC) technique is less sensitive and often more difficult to interpret in antibody identification. In some cases, however, this method was the only one that gave positive reactions. In 214 instances antibodies were identified by the AA techniques, about half of which were anti-D's, the others being RBC antibodies of more or less rare varieties. The relative frequency of these rare antibodies was higher in the group of manual-negative sera, 47 versus 21%, normally found. Positive identification but no clear-cut blood type antibody was found in 155 cases (19%). Indications were obtained that some of the reactions were due to HL-A antibodies. Antibodies to HL-A7 and HL-A5 were found to react with red cells. In addition the AA techniques seemed to be more sensitive to detect RBC autoantibodies than standard manual methods. However, the usefulness of the AA techniques for routine antibody identification is diminished by the relative slowness of the machine.