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ABSTRACT: Parietal cell antibody is a marker for autoimmune gastritis. With identification of gastric H/K ATPase as its molecular target, ELISAs have been introduced. We compared performance of ELISA with immunofluorescence in a retrospective and prospective sera set and correlated the results with intrinsic factor antibody. In 138 retrospective sera selected for positivity or negativity for intrinsic factor antibody, 87 reacted with gastric H/K ATPase by Euroimm ELISA but only 62 reacted by immunofluorescencence.. Similar results were obtained with Inova ELISA with 78 positives that were also positive by Euroimm ELISA. In 161 prospective sera, 29 sera tested positive by ELISA compared to 24 by immunofluorescence. ELISA positive but immunofluoresnce negative sera are bona fide positives because a representative set of 16 sera reacted with both 95kD α and 60-90kDβ subunits of gastric H/K ATPase. ELISA values rose with age regardless of whether immunofluorescence tests were positive or negative. Of 53 sera containing antibody to intrinsic factor, 46/53 (87%) reacted to gastric H/K ATPase by ELISA. Taken together, the data indicates an enhanced detection rate by ELISA over immunofluorescence and validates it as a robust diagnostic assay for parietal cell antibody. As parietal cell antibody marks asymptomatic autoimmune gastritis that may progress to end stage gastric atrophy and haematological complications, and as autoimmune gastritis is associated with autoimmune thyroiditic and type 1 diabetes mellitus, early detection of parietal cell antibody by a sensitive ELISA will enable early follow-up of at risk subjects.
Autoimmunity 07/2012; 45(7):527-32. · 2.47 Impact Factor
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ABSTRACT: The Australasian Maternity Outcomes Surveillance System (AMOSS) conducts surveillance and research of rare and serious conditions in pregnancy. This multi-centre population health study is considered low risk with minimal ethical impact.
To describe the ethics/governance review pathway undertaken by AMOSS.
Prospective, descriptive study during 2009-2011 of the governance/ethical review processes required to gain approval for Australian and New Zealand (ANZ) maternity units with more than 50 births per year (n = 303) to participate in AMOSS.
Review processes ranged from a single application for 24 NZ sites, a single application for eligible hospitals in two Australian states, full Health Research Ethics Committee (HREC) applications for individual hospitals, through simple letters of support. As of September 2011, 46 full/expedited ethics applications, 131 site governance applications and 136 letters of support requests were made over 33 months, involving an estimated 3261 hours by AMOSS staff/investigators, and an associated resource burden by participating sites, to obtain approval to receive nonidentifiable data from 291 hospitals.
The AMOSS research system provides an important resource to enhance knowledge of conditions that cause rare and serious maternal morbidity. Yet the highly variable ethical approval processes required to implement this study have been excessively repetitive and burdensome. This process jeopardises timely, efficient research project implementation, without corresponding benefits to research participants. The resource burden to establish research governance for AMOSS confirms the urgent need for the Harmonisation of Multi-centre Ethical Review (HoMER) to further streamline ethics/governance review processes for multi-centre research.
Australian and New Zealand Journal of Obstetrics and Gynaecology 12/2011; 52(2):195-203. · 1.24 Impact Factor
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ABSTRACT: To compare smooth muscle antibody (SMA) patterns in tissue sections with patterns in an immunofluorescence assay (IFA) using a rat intestinal epithelial cell line and results from an F-actin IgG ELISA.
SMA positive sera (n = 188) were classified by immunofluorescence staining of rodent kidney, stomach and liver sections as SMA-T (tubules) (n = 124) or SMA-V (vessels) (n = 64). The F-actin pattern on the rat epithelial cell line was identified by immunofluorescence staining of actin cables that was confirmed by dual immunofluorescence co-localisation with phalloidin.
Of 124 SMA-T positive sera, 123 reacted with the epithelial cell line and 120 with F-actin by ELISA, giving sensitivity for detection of anti-F-actin antibody of 99% and 97%, respectively. Of 64 SMA-V positive sera, four reacted with the epithelial cell line (6%) and 41 with F-actin by ELISA (64%). Tests of 493 normal blood donors and 100 disease controls yielded specificities of 584/593 (98.5%) and 562/593 (94.8%) for the cell line IFA and F-actin ELISA, respectively.
The rat epithelial cell line IFA is a robust diagnostic assay for anti-F-actin antibody that can either replace the routine screening for actin-reactive SMA-T antibody in tissue sections or be used as a confirmatory assay for anti-F-actin antibody after screening by F-actin ELISA or on rodent tissue sections by IFA.
Pathology 01/2010; 42(5):463-9. · 2.38 Impact Factor
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ABSTRACT: Through the Australian and New Zealand Haemostasis Registry, we report on the Australian and New Zealand experience with recombinant activated factor VII (rFVIIa) in obstetric patients.
The role of rFVIIa for off-label indications, including trauma, cardiac surgery, and severe postpartum hemorrhage, remains controversial. The Haemostasis Registry established by Monash University in Melbourne, Australia monitors off-label use of rFVIIa across Australia and New Zealand. The purpose of this study was to summarize Registry data for all obstetric hemorrhage patients treated with rFVIIa at participating hospitals between January 2002 and July 2008. The primary outcome measures were reduction or cessation of bleeding (positive therapeutic response), mortality, and hysterectomy rate.
During the study period, the Registry received data for 2128 patients. This included 110 cases of administration of rFVIIa in obstetric patients from 38 hospitals, comprising 5% of the total Registry population, 105 of whom were treated for acute hemorrhage. Women received median (interquartile range) individual doses of 92 microg/kg (73-100) of rFVIIa (median total dose 92 microg/kg [58-108]), and 78% of patients received a single dose. The positive response rate to rFVIIa was 76% with 64% responding to the first dose. Ninety-one percent of women were alive at 28 days. Forty-three women (41%) underwent hysterectomy before receiving rFVIIa and, of those remaining, 13 (21%) required hysterectomy after rFVIIa therapy. Two thromboembolic events (1 pulmonary embolism and 1 deep venous thrombosis) and 1 case of hypoxic-ischemic encephalopathy resulting from severe anoxia were reported.
The reported effect of rFVIIa in many, but not all, obstetric cases was positive. There was no mortality as a result of thromboembolic complications. Randomized, controlled trials are required to confirm its safety and efficacy and to assess the possibility that use at an earlier stage in treatment of severe postpartum hemorrhage may avoid the need to resort to postpartum hysterectomy for control of bleeding, thus preserving fertility.
Anesthesia and analgesia 12/2009; 109(6):1908-15. · 3.08 Impact Factor
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ABSTRACT: Most laboratories screen for antineutrophil cytoplasmic antibodies (ANCA) with indirect immunofluorescence (IIF) and confirm cytoplasmic (C-ANCA) and perinuclear (P-ANCA) staining with ELISAs for proteinase 3 (PR3) and myeloperoxidase (MPO) specificities. This study determined the concordance of ANCA test results from 48 diagnostic laboratories participating in a national Quality Assurance programme, that used different assays and methods and varied in expertise. Laboratories were circulated with a questionnaire about their techniques, and provided with 24 sera for testing over a 30 month period. Results for individual sera were compared with the 'observed consensus' found in more than 50% of laboratories. The 23 laboratories (48%) that responded to the questionnaire used 5 different IIF substrates and 11 ELISAs, and differed in other aspects of testing. Concordance for ANCA test results was greater for IIF-positive (n=22, median 96%, range 68%-100%) than an IIF-negative serum (median 64%); for C-ANCA (n=8, median 89%, range 66-100%) rather than P-ANCA (n=10, median 76%, range 52-88%); for MPO-ANCA (n=5, median 100%) rather than PR3-ANCA (n=7, median 89%, range 82-100%); and for strongly-positive (n=2, median 97%, range 96-97%) rather than low positive PR3-ANCA (n=4, median 80%, range 74-86%). Concordance for test results might be improved with further standardisation of testing methodologies.
Journal of immunological methods 06/2009; 347(1-2):19-23. · 2.35 Impact Factor
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ABSTRACT: Antineutrophil cytoplasmic antibodies (ANCA) levels have been thought to follow disease activity, with levels being high at presentation, declining with treatment and increasing just before relapse. However, we have shown that ANCA often persist for many years in patients with clinically inactive Wegener's granulomatosis. ANCA assays are less sensitive for treated disease than for active disease, and the levels in treated patients produce different results in different assay systems. ANCA often persist for years without relapse, and the risk of relapse probably depend on levels that are critical for any individual patient. The capture enzyme-linked immunosorbent assays may be more sensitive in detecting early relapse. Relapse is more common when ANCA levels are high but, although elevated, ANCA levels are lower in relapse than at presentation. Standardized ANCA levels for the definitions of remission and relapse may not be possible, and the optimal ANCA testing protocol for treated disease remains unclear.
APMIS. Supplementum 06/2009;
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ABSTRACT: Smooth muscle antibody (SMA) with F-actin reactivity has been reported as a diagnostic marker of autoimmune hepatitis. We re-visited this relationship by randomly selecting SMA-positive sera to test for reactivity with F-actin by ELISA. We correlated such reactivity with liver function tests.
Sera positive for SMA by indirect immunofluorescence were tested for reactivity by F-actin ELISA and the results correlated with liver function tests.
89 SMA-positive sera reacted with F-actin by ELISA. Of these, 35 (39%) had normal liver enzymes, while 54 (60%) had elevated liver enzymes. There was no difference between the groups with respect to age at presentation, female preponderance or presence of anti-nuclear antibody. In both groups, high titre SMA antibody predominantly with immunofluorescence staining of renal glomeruli and peritubular fibrils of renal tubules ('G/T' subset) correlated with makedly elevated F-actin values by ELISA. 'Actin cables' by immunofluorescence staining of Hep-2 cells were infrequently found in both groups.
This is the first report of SMA with F-actin reactivity in subjects with normal liver function.
Pathology 01/2009; 41(6):572-5. · 2.38 Impact Factor
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ABSTRACT: This study evaluated the usefulness of testing sera with perinuclear antineutrophil cytoplasmic antibodies (P-ANCA) on formalin-fixed neutrophils to differentiate between vasculitis and other diseases, and to distinguish between myeloperoxidase (MPO) and other antigen targets. Sera from active (n=24) or treated vasculitis (n=23) and non-vasculitic disease (n=37) were tested in 3 ethanol- and 2 formalin-fixed neutrophil assays, and in 12 MPO-ANCA ELISAs. The sensitivity of demonstrating that P-ANCA became cytoplasmic on formalin-fixed cells in vasculitis, and negative in non-vasculitic disease was 75-79% in different assays compared with 88% for positivity in the majority of MPO-ANCA ELISAs. The sensitivity and specificity of demonstrating that P-ANCA became cytoplasmic where the target was MPO, and disappeared with other antigens were 89% and 74-80% respectively in different assays. P-ANCA specificity for MPO should be confirmed in one of the better-performing MPO-ANCA ELISAs.
Journal of Immunological Methods 10/2008; 339(2):141-5. · 2.20 Impact Factor
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ABSTRACT: This study compared the performance of a flow cytometric immunoassay for antineutrophil cytoplasmic antibodies (ANCA) directed against proteinase 3 (PR3) and myeloperoxidase (MPO), with indirect immunofluorescence (IIF) and ELISAs from 12 different manufacturers. Sera were from patients with active (n=55) or treated (n=68) small vessel vasculitis, or inflammatory bowel disease (IBD, n=22). The immunoassay specificity was 88% compared with 96% for IIF and 94% (median, range 91-96%) for both ELISAs. Its sensitivity in treated disease was 82% compared with 84% for IIF and 69% (median, range 57-82%) for the ELISAs. The immunoassay's specificity was 88% which was the same as the median for both ELISAs (range 84-95%). The PR3- and MPO-ANCA immunoassay was almost as sensitive as IIF, and more sensitive than, but just as specific as, most ELISAs, in detecting ANCA in active and treated vasculitis. A major advantage of this assay is its ability to be further modified to simultaneously screen for a panel of autoantibodies relevant to vasculitis.
Journal of Immunological Methods 08/2008; 336(2):104-12. · 2.20 Impact Factor
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ABSTRACT: Maternal mortality has traditionally been the key element in the monitoring of maternal health and adequacy of obstetric services in Australia and around the world. In developed countries, the ability of maternal mortality to serve this purpose is reduced because of the rarity of maternal mortality, reflected in very low maternal mortality ratios. Internationally, there has been increasing interest in severe maternal morbidity as an indicator to monitor maternal health and maternity services. The aim of this paper is to critically examine the capacity to measure and monitor maternal morbidity in Australia. There is a paucity of reliable maternal morbidity data in Australia; Australia is lagging behind peer countries that are endeavouring to monitor severe maternal morbidity. Dedicated efforts and adequate resources are needed in order to monitor severe maternal morbidity in Australia.
Australian and New Zealand Journal of Obstetrics and Gynaecology 03/2008; 48(1):17-25. · 1.24 Impact Factor
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ABSTRACT: This study evaluated the performance of 12 assays for antineutrophil cytoplasmic antibodies (ANCA) directed against proteinase 3 (PR3) and myeloperoxidase (MPO) in 55 active and 68 treated cases of vasculitis and in nonvasculitic disease. It examined within- and between-assay precision; binding curves, binding levels, and interassay consistency; and sensitivity, specificity, and receiver operating characteristic analysis. All assays were highly sensitive for active vasculitis (median, 94%; range, 91%-96%), but sensitivities were more varied in treated disease (median, 69%; range, 57%-82%). Binding curves and binding levels were also very variable in PR3-ANCA and MPO-ANCA assays. This has implications for studies correlating ANCA levels with disease activity and in developing ANCA-based treatment guidelines. PR3-ANCA and MPO-ANCA assays need to be standardized as a matter of urgency, but in the meantime, individual laboratories must understand the limitations of the assays used, especially with low-level ANCA in treated vasculitis and nonvasculitic disease.
American Journal of Clinical Pathology 02/2008; 129(1):42-53. · 2.60 Impact Factor
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ABSTRACT: Antineutrophil cytoplasmic antibodies (ANCA) are autoantibodies directed against antigens found in the cytoplasmic granules of neutrophils and monocytes. ANCA testing is usually performed to help diagnose or exclude Wegener's granulomatosis and microscopic polyangiitis. The three most commonly used assays are indirect immunofluorescence (IIF) and the direct and 'capture' enzyme-linked immunosorbent assays (ELISAs) for ANCA directed against proteinase 3 (PR3) and myeloperoxidase (MPO). Although the International Consensus Statement for Testing and Reporting ANCA recommends that all sera are screened for ANCA by IIF and that IIF-positivity is confirmed by direct ELISAs, some laboratories test by direct ELISA alone, others screen with direct ELISA and confirm positive sera by IIF, and a few use capture ELISAs. This chapter discusses the various forms of vasculitis associated with ANCA, the usefulness of each of the ANCA assays and how ANCA testing can be used in the management of patients with small-vessel vasculitis.
Bailliè re s Best Practice and Research in Clinical Rheumatology 05/2005; 19(2):263-76. · 2.65 Impact Factor
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ABSTRACT: To investigate differences in anti-cardiolipin antibody (aCL) testing and reporting practices among diagnostic laboratories participating in the RCPA Quality Assurance Program (QAP) Immunology Program.
A survey was sent to all 58 laboratories enrolled for aCL testing in the RCPA QAP Program requesting the following information: (1). manufacturer/type of assay; (2). isotype tested; (3). use of calibrators and controls; (4). whether calibrators, control and patient specimens were performed in singles or duplicates; (5). whether results were reported semi-quantitatively and/or numerically; (6). values used to define negative/positive and semi-quantitative cut-offs and how they were determined; and (7). whether interpretative comments were provided and their content.
Thirty-two surveys (55%) were received. Significant differences were present particularly in the following areas: (1). whether IgM isotype aCL testing was performed routinely or only on specific request; (2). whether controls/calibrators/specimens were tested in singles or duplicates; (3). whether results were reported numerically and/or semi-quantitatively; (4). the values used to define negative/positive and semi-quantitative range cut-offs; and (5). whether interpretative comments were provided and their content.
These differences in testing and reporting practices are likely to contribute to the variation in aCL results reported by different laboratories, particularly among those using the same assay.
Pathology 05/2004; 36(2):174-81. · 2.38 Impact Factor
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Richard Wong,
Emmanuel Favaloro, Wendy Pollock,
Robert Wilson,
Michelle Hendle,
Stephen Adelstein,
Karl Baumgart,
Paul Homes,
Stuart Smith,
Richard Steele,
Allan Sturgess,
David Gillis
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ABSTRACT: To assess the intra-assay (intra-run) and inter-assay (inter-run) variation of commercial and in-house IgG and IgM anti-cardiolipin antibody (aCL) assays/kits, and to determine an appropriate maximum value for inclusion in consensus guidelines.
Frozen aliquots of two patient specimens and one commercial control were sent to nine laboratories for the evaluation of eight commercial kits and one in-house assay. Intra-assay and inter-assay evaluations were performed with all three samples for IgG aCL, and one patient specimen for IgM aCL.
The IgG and IgM aCL values varied considerably between the nine assays/kits. The majority of assays/kits demonstrated less than 20% intra-assay and inter-assay variation, with lower intra-assay and inter-assay variation observed with the commercial control. Single calibrator assays were not consistently associated with higher inter-assay variation than multi-point calibrator assays.
An inter-assay coefficient of variation of 20% was determined to be an appropriate maximum value for inclusion in the Australasian aCL Working Party consensus guidelines. Improved standardisation between different assay/kits is still required.
Pathology 05/2004; 36(2):182-92. · 2.38 Impact Factor
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Richard C W Wong,
David Gillis,
Stephen Adelstein,
Karl Baumgart,
Emmanuel J Favaloro,
Michelle J Hendle,
Paul Homes, Wendy Pollock,
Stewart Smith,
Richard H Steele,
Allan Sturgess,
Robert J Wilson
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ABSTRACT: Consensus guidelines on anti-cardiolipin antibody (aCL) testing have been developed to help minimise laboratory variation in the performance and reporting of aCL assays. These guidelines include minimum, optimum and optional recommendations for the following aspects of aCL testing and reporting: (1) isotype of aCL tested; (2) specimen type; (3) controls and assay precision; (4) calibrators; (5) patient samples; (6) rheumatoid factors and IgM aCL testing; (7) reporting of results; (8) cut-off values; and (9) interpretative comments. Abbreviations: aCL, anti-cardiolipin antibodies; APS, anti-phospholipid antibody syndrome; ASCIA, Australasian Society of Clinical Immunology and Allergy; ASTH, Australasian Society of Thrombosis and Haemostasis; beta2-GPI=beta2-glycoprotein I; ELISA, enzyme-linked immunosorbent assay; NCCLS, National Committee for Clinical Laboratory Standards; HSANZ, Haematology Society of Australia and New Zealand; QAP, Quality Assurance Program; RCPA, Royal College of Pathologists of Australasia; %CV, inter-assay inter-run coefficient of variation.
Pathology 03/2004; 36(1):63-8. · 2.38 Impact Factor
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Judy Savige,
Wayne Dimech,
Marvin Fritzler,
James Goeken,
E Chris Hagen,
J Charles Jennette,
Rob McEvoy,
Charles Pusey, Wendy Pollock,
Michelle Trevisin,
Allan Wiik,
Richard Wong
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ABSTRACT: Antineutrophil cytoplasmic antibody (ANCA) tests are used to diagnose and monitor inflammatory activity in Wegener granulomatosis, microscopic polyangiitis and its renal-limited variant (pauci-immune crescentic glomerulonephritis), and Churg-Strauss syndrome. The International Consensus Statement on testing and reporting of ANCA states that ANCA are demonstrated most readily in these conditions by using a combination of indirect immunofluorescence (IIF) of normal peripheral blood neutrophils and enzyme-linked immunosorbent assays (ELISAs) that detect ANCA specific for proteinase 3 or myeloperoxidase. The group that produced the International Consensus Statement has developed guidelines for the corresponding quality control activities, examples of comments for various IIF patterns and ELISA results, and recommendations for ANCA testing when inflammatory bowel disease and other nonvasculitic ANCA-associated autoimmune diseases are suspected.
American Journal of Clinical Pathology 10/2003; 120(3):312-8. · 2.60 Impact Factor
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ABSTRACT: There has been little research conducted on critically ill pregnant and postnatal women. When developing a research protocol to conduct a prospective multi-centre survey on this study population, we found there were vital concerns that needed addressing prior to the research proceeding. Prompt identification of the study population and valid, reliable data collection were two aspects that needed particular attention with study recruitment potentially occurring in a total of eleven clinical areas from seven hospitals. In this paper we outline the particular issues faced by us when conducting multi-centre research on a study population that occurs infrequently and unpredictably, and when there is a necessary urgency to identify eligible study participants. The key strategy to overcome these difficulties, was the creation of 'Core Research Teams' in each clinical area. Our experience of using Core Research Teams in our pilot study is described in this paper. We found that the Core Research Team model is a very positive strategy to overcome the methodological challenges when operating a multi-centre study.
Contemporary nurse: a journal for the Australian nursing profession 01/2003; 14(1):95-105. · 0.67 Impact Factor
