scholarly journals Performance Characteristics of the PolyTiter Immunofluorescent Titration System for Determination of Antinuclear Antibody Endpoint Dilution

2002 ◽  
Vol 9 (2) ◽  
pp. 329-332 ◽  
Author(s):  
Karen A. Flessland ◽  
Helen R. Landicho ◽  
Kimberlee K. Borden ◽  
Harry E. Prince

ABSTRACT Conventional screening for circulating antinuclear antibodies (ANA) is generally performed by immunofluorescent (IF) microscopy with a 1:40 dilution of serum. Intensity of IF staining is then semiquantitated by using twofold serial dilutions, where the highest dilution in which staining intensity equals the endpoint control is expressed as an endpoint titer. The PolyTiter Immunofluorescent Titration system (Polymedco, Inc.) facilitates ANA-IF assay (IFA) testing by relating the intensity of IF staining to reference calibrators (defined in PolyTiter units), providing an endpoint titer directly from a 1:40 dilution. This study was conducted to assess the performance characteristics of the PolyTiter system. Two technologists each evaluated 10 replicates of three specimens and two controls on five sequential days. Endpoint dilution agreement (defined as ±2 dilutions) with the reference was 100% for all controls and for all specimens by one technologist. The second reader reported agreement of 98, 88, and 100% for the low, medium, and high specimens, respectively. Analysis of PolyTiter unit values yielded between-reader, between-run, and within-run precision coefficients of variation of less than 10%. The variance component in the lot-to-lot analysis was zero, indicating all of the variation was due to run-to-run differences. Overall endpoint dilution agreement between PolyTiter and serial dilution in the evaluation of 125 specimens at three sites was 90, 93, and 86%. Pattern identification with the PolyTiter was similar to that with serial dilution. The PolyTiter system demonstrates acceptable performance for routine ANA-IFA testing in the clinical laboratory.

1984 ◽  
Vol 30 (7) ◽  
pp. 1227-1228 ◽  
Author(s):  
N W Tietz ◽  
D F Shuey

Abstract On evaluation, an S-type alpha-amylase inhibitor from Sigma shows essentially the same performance characteristics as the inhibitor from wheat germ prepared according to O'Donnell and McGeeney (Biochim Biophys Acta 422:159, 1976). This inhibitor is now commercially available; thus any clinical laboratory can perform isoamylase determinations with a simple, fast procedure that is also suitable for emergency situations.


Author(s):  
Ranu Kumar ◽  
Prasad Kapildeo

We are traditionally used Microscope in clinical laboratory for determination of white blood cells of human blood smear. Now, in this study we were used Foldscope with Smartphone in the place of Microscope and examine many samples of human blood smear which was collected from local diagnostic centers. We were very easily quantity & morphology analysis of all types of WBC cells such as Neutrophils, Lymphocytes, Monocytes, Eosionophils, Basophils in blood smear with the help of Foldscope & image taken by Smartphone. The main objective of this study is to use Foldscope for quantity & morphology analysis of human WBCs at field level especially poor resource area where healthcare services or centers is not available & where carry of microscope is not possible.


2020 ◽  
Vol 58 (6) ◽  
pp. 1002-1009 ◽  
Author(s):  
Caroline Le Goff ◽  
Jordi Farre-Segura ◽  
Violeta Stojkovic ◽  
Patrice Dufour ◽  
Stéphanie Peeters ◽  
...  

AbstractHistorically, the determination of low concentration analytes was initially made possible by the development of rapid and easy-to-perform immunoassays (IAs). Unfortunately, typical problems inherent to IA technologies rapidly appeared (e.g. elevated cost, cross-reactivity, lot-to-lot variability, etc.). In turn, liquid chromatography tandem mass spectrometry (LC-MS/MS) methods are sensitive and specific enough for such analyses. Therefore, they would seem to be the most promising candidates to replace IAs. There are two main choices when implementing a new LC-MS/MS method in a clinical laboratory: (1) Developing an in-house method or (2) purchasing ready-to-use kits. In this paper, we discuss some of the respective advantages, disadvantages and mandatory requirements of each choice. Additionally, we also share our experiences when developing an in-house method for cortisol determination and the implementation of an “ready-to-use” (RTU) kit for steroids analysis.


2018 ◽  
Vol 76 (2) ◽  
pp. 442-451 ◽  
Author(s):  
Gaya Gnanalingam ◽  
Mark J Butler ◽  
Thomas R Matthews ◽  
Emily Hutchinson ◽  
Raouf Kilada

Abstract In crustaceans, ecdysis was long believed to result in the loss and replacement of all calcified structures, precluding the use of conventional ageing methods. However, the discovery of bands in the gastric ossicles of several crustaceans with some correlation with age suggests that direct age estimation may be possible. We applied this method to a tropical spiny lobster, Panulirus argus, one of the most iconic and economically valuable species in the Caribbean. The presence of growth bands was investigated using wild lobsters of unknown age and was validated with captive reared lobsters of known age (1.5–10 years) from the Florida Keys, Florida (USA). Bands were consistently identified in ptero- and zygo-cardiac ossicles of the gastric mill and did not appear to be associated with moulting. Validation with known age animals confirms that bands form annually. Counts between independent readers were reproducible with coefficients of variation ranging from 11% to 26% depending on reader experience and the structure used. This study demonstrates, for the first time, that direct age determination of P. argus is possible.


1983 ◽  
Vol 104 (1) ◽  
pp. 1-5 ◽  
Author(s):  
J. Leppäluoto ◽  
L. Rönnberg ◽  
P. Ylöstalo

Abstract. Seven patients suffering from severe endometriosis were treated with danazol 200 mg × 3 daily for 6 months. Clinical symptoms were alleviated and menses disappeared in response to the treatment. After cessation of the treatment the menstrual bleedings returned in 1–3 months. Blood samples for determination of gonadotrophins, prolactin (Prl), oestradiol (E2), progesterone, thyroid hormones and thyrotrophin in radioimmunoassays were taken and a combined TRF and LRF test carried out in the follicular phase before treatment, at the 6th month of treatment and after reappearance of the first menses. There were no statistically significant changes in the basal levels of serum FSH, LH or TSH during the danazol treatment. Neither was there any change in episodic secretions of FSH, LH or Prl, as determined by the mean coefficients of variation of the hormone levels in seven consecutive samples taken at 20 min intervals. On the other hand, serum E2, Prl and thyroid hormone levels were significantly decreased in the 6th month of treatment. In the TRF-LRF test the responses of serum FSH and LH were significantly higher and those of serum Prl and TSH significantly lower during danazol treatment than before. Prl responses remained lowered after the treatment. It appears that low serum oestrogen levels, induced by the danazol treatment, sensitize the pituitary gonadotrophs to exogenous LRF, but make the sensitivity of thyrotrophs and lactotrophs lower to exogenous TRF. These results thus indicate that danazol does not make the pituitary gonadotrophs insensitive to LRF, but danazol may rather inhibit the secretion of hypothalamic LRF.


2020 ◽  
Vol 58 (9) ◽  
pp. 1489-1497 ◽  
Author(s):  
Lisa K. Peterson ◽  
Anne E. Tebo ◽  
Mark H. Wener ◽  
Susan S. Copple ◽  
Marvin J. Fritzler

AbstractBackgroundThe indirect immunofluorescence assay (IFA) using HEp-2 cell substrates is the preferred method by some for detecting antinuclear antibodies (ANA) as it demonstrates a number of characteristic staining patterns that reflect the cellular components bound as well as semi-quantitative results. Lack of harmonized nomenclature for HEp-2 IFA patterns, subjectivity in interpretation and variability in the number of patterns reported by different laboratories pose significant harmonization challenges. The main objectives of this study were to assess current practice in laboratory assessment of HEp-2 IFA, identify gaps and define strategies to improve reading, interpretation and reporting.MethodsWe developed and administered a 24-item survey based on four domains: educational and professional background of participants, current practice of HEp-2 IFA testing and training, gap assessment and the perceived value of International Consensus on Antinuclear Antibody Patterns (ICAP) and other factors in HEp-2 IFA assessment. The Association of Medical Laboratory Immunologists (AMLI) and American Society for Clinical Pathology administered the survey from April 1 to June 30, 2018, to members involved in ANA testing. This report summarizes the survey results and discussion from a dry workshop held during the 2019 AMLI annual meeting.ResultsOne hundred and seventy-nine (n = 179) responses were obtained where a significant number were clinical laboratory scientists (46%), laboratory directors (24%), supervisors (13%) or others (17%). A majority of respondents agreed on the need to standardize nomenclature and reporting of HEp-2 IFA results. About 55% were aware of the ICAP initiative; however, among those aware, a significant majority thought its guidance on HEp-2 IFA nomenclature and reporting is of value to clinical laboratories. To improve ICAP awareness and further enhance HEp-2 IFA assessment, increased collaboration between ICAP and the clinical laboratory community was suggested with emphasis on education and availability of reference materials.ConclusionsBased on these suggestions, future efforts to optimize HEp-2 IFA reading, interpretation and reporting would benefit from more hands-on training of laboratory personnel as well as continuous collaboration between professional organizations, in vitro diagnostic manufacturers and clinical laboratories.


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