The measurement of luteinising hormone in the western grey kangaroo (Macropus fuliginosus ocydromus) and the black-flanked rock wallaby (Petrogale lateralis lateralis)

2009 ◽  
Vol 31 (1) ◽  
pp. 61 ◽  
Author(s):  
Phillip Matson ◽  
Christopher Mayberry ◽  
Nicole Willers ◽  
Margaret A. Blackberry ◽  
Graeme B. Martin
Keyword(s):  
Enzyme Immunoassay ◽  
Luteinising Hormone ◽  
Standard Curve ◽  
Grey Kangaroo ◽  
Western Grey Kangaroo

An enzyme immunoassay with an anti-bovine-LH antibody (518B7) was applied to female western grey kanagaroos (Macropus fuliginosus ocydromus) and black-flanked rock wallabies (Petrogale lateralis lateralis). Validation showed parallelism to the assay standard curve, and significant increases in plasma LH concentrations after challenging animals with intramuscular GnRH.

276. The technical and biological validation of an LH assay for use with the Western Grey Kangaroo (Macropus fuliginosis) and Black-flanked Rock Wallaby (Petrogale lateralis lateralis)

2008 ◽  
Vol 20 (9) ◽  
pp. 76
Author(s):  
P. Matson ◽  
C. Mayberry ◽  
N. Willers ◽  
M. A. Blackberry ◽  
G. B. Martin
Keyword(s):  
Plasma Concentrations ◽  
Standard Curve ◽  
Biological Validation ◽  
Grey Kangaroo ◽  
Coefficients Of Variation ◽  
Western Grey Kangaroo ◽  
Technical Validation ◽  
Zoo Biology ◽  
Smithsonian Institute ◽  
And Control

Methods for the measurement of marsupial LH invariably rely upon the similarity of the LH molecule between different species and usually use anti-ovine or anti-bovine LH antibody and an ovine or bovine labelled LH preparation. Initial attempts to measure plasma LH in the Western Grey Kangaroo with assays using antibodies to 4 different isoforms of ovine LH raised in 7 different rabbits were unsuccessful. An enzymeimmunoassay (EIA) developed for the Asian elephant (Zoo Biology 23:45–63) was then applied to the Western Grey Kangaroo and the Black-flanked Rock Wallaby. This EIA has an anti-bovine-LH monoclonal antibody (518B7 provided by Dr Jan Roser, University of California, Davis, USA), biotinylated ovine LH label and bovine LH standard (NIADDK-oLH-26 and NIH-bLH-B10, both provided by Dr Janine Brown and Nicole Abbondanza, Smithsonian Institute, Front Royal, Virginia USA). Technical validation showed that serial dilution down to 1:8 of plasma from 7 individuals of each species showed parallelism to the assay standard curve, and control samples (1.24–5.30 ng/mL) had between-assay coefficients of variation <9%. Biological validation was achieved by challenging animals with intramuscular GnRH (Fertagyl®, 2.5 µg/kg) and measuring LH before and 25 min after the injection. Significant increases in plasma concentrations of LH (mean ± sem; all P > 0.0005) were seen after GnRH for both the Western Grey Kangaroo (from 5.0 ± 0.8 ng/mL to 9.4 ± 1.2 ng/mL; n = 19) and the Black-flanked Rock Wallaby (from 6.0 ± 0.7 ng/mL to 10.6 ± 0.6 ng/mL; n = 28). In conclusion, this assay can be successfully used to measure LH in these two species.

Evaluation of a solid-phase enzyme immunoassay for human choriogonadotropin beta subunit.

1983 ◽  
Vol 29 (11) ◽  
pp. 1964-1966 ◽  
Author(s):  
D D Davey ◽  
M M Sample ◽  
T O Oei
Keyword(s):  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Cross Reactivity ◽  
Beta Subunit ◽  
Neoplastic Disease ◽  
Standard Curve ◽  
Human Choriogonadotropin ◽  
Beta Hcg ◽  
Substantial Problem ◽  
Sample Dilution

Abstract We evaluated a quantitative solid-phase enzyme immunoassay for human choriogonadotropin beta subunit (beta-HCG) with anti-beta-HCG:horseradish peroxidase conjugate, recently marketed by Abbott Laboratories. We compared results on 56 patients' serum specimens, obtained mostly for followup of neoplastic disease, with those by a competitive radioimmunoassay kit. The correlation was good, the differences being of little clinical significance. Linear regression in the low and intermediate ranges gave a slope of 0.93, a y-intercept of 0.34, and a correlation coefficient of 0.97. Precision studies yielded an interassay CV of 6.4% in the intermediate range and 13% in the low range. Sensitivity was 0.69 int. unit/L. Cross reactivity was 1 to 2% with specimens fortified with lutropin or follitropin. The only substantial problem was with linearity in the upper part of the standard curve, especially in the interval, 100-200 int. units/L. This problem is obviated by adequate sample dilution.

Fluoroacetate Tolerance, a Genetic Marker in Some Australian Mammals.

1979 ◽  
Vol 27 (3) ◽  
pp. 363 ◽  
Author(s):  
AJ Oliver ◽  
DR King ◽  
RJ Mead
Keyword(s):  
Genetic Marker ◽  
Geographic Origin ◽  
Tammar Wallaby ◽  
Southeastern Australia ◽  
Toxic Plants ◽  
Grey Kangaroo ◽  
Western Grey Kangaroo ◽  
History Of ◽  
Genetic Tolerance ◽  
High Level

The toxin fluoroacetate occurs naturally in many southwestern Australian species of the legume genera Gastrolobium and Oxylobium. No fluoroacetate-bearing species are known from southeastern Australia. Herbivores have evolved a high level of genetic tolerance to this toxin; this has persisted in some mammalian herbivores whose range now extends beyond the range of the toxic plants. Other species of mammals have acquired tolerance since extending their range into south-western Australia. This tolerance can be used as a genetic marker to identify the geographic origin and trace the subsequent spread of herbivorous mammals in southern Australia. In this paper, this marker has been used to clarify the recent evolutionary history of the western grey kangaroo, the tammar wallaby and the bush rat.

scholarly journals Development of enzyme immunoassay for endogenous ouabain-like compound in human plasma

1997 ◽  
Vol 43 (5) ◽  
pp. 715-722 ◽  
Author(s):  
Steven Harwood ◽  
John A Little ◽  
Gerard Gallacher ◽  
David Perrett ◽  
Raymond Edwards ◽  
...  
Keyword(s):  
Detection Limit ◽  
Healthy Volunteers ◽  
Enzyme Immunoassay ◽  
Human Plasma ◽  
Hplc Analysis ◽  
Cross Reactivity ◽  
Standard Curve ◽  
Endogenous Ouabain ◽  
Endogenous Steroid ◽  
Hplc Study

Abstract Widespread evidence supports the existence of an endogenous digitalis-like compound in mammals. We report here the development of a novel enzyme immunoassay for ouabain that, in conjunction with a detailed HPLC study, identifies a ouabain-like compound (OLC) in extracted human plasma. The assay is sensitive—minimum detection limit for OLC 37 pmol/L (11 pmol/L in plasma)—and has a working range (between-assay CV &lt;10%) of 180–10 000 pmol/L (54–3000 pmol/L in plasma). Mean recoveries of ouabain added to plasma ranged from 90% to 100%, and plasma extracts diluted in parallel to the standard curve. Plasma OLC concentrations in 10 healthy volunteers averaged 92 pmol/L (range 55–168), assuming 100% cross-reactivity of OLC in the ouabain assay. HPLC analysis with two distinct chromatographic conditions demonstrated that endogenous human plasma OLC co-eluted with authentic ouabain. The enzyme immunoassay is rapid and easy to perform and will support further investigation of the nature of this controversial endogenous steroid.

Sandwich enzyme immunoassay of osteocalcin in serum with use of an antibody against human osteocalcin

1993 ◽  
Vol 39 (6) ◽  
pp. 942-947 ◽  
Author(s):  
D A Monaghan ◽  
M J Power ◽  
P F Fottrell
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Sandwich Elisa ◽  
Normal Subjects ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Standard Curve ◽  
Microtiter Plates ◽  
Human Osteocalcin

Abstract We have developed and thoroughly validated a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA) on microtiter plates for osteocalcin in human serum with use of an antibody raised against human osteocalcin. We used a monoclonal antibody against bovine osteocalcin as the capture antibody; the second antibody was a polyclonal antibody against human osteocalcin. The amount of bound second antibody was determined with use of swine anti-rabbit antibody labeled with horseradish peroxidase. We demonstrated independence of volume and determined the recovery of added standard and within- and between-assay precision. The minimal detection limit for osteocalcin was between 1.0 and 1.5 micrograms/L and the midpoint of the standard curve ranged from 14 to 17 micrograms/L. The intraassay CV was &lt; or = 8% in the range 2.7-52 micrograms/L; the interassay CV was usually &lt; or = 15% in the same range. Analytical recovery of human osteocalcin standard added to serum samples was consistently &gt; 90%. Values for osteocalcin measured in serum from 44 normal subjects were similar to those obtained with a competitive enzyme immunoassay (EIA) that used a monoclonal antibody against bovine osteocalcin. There was a good correlation between the two assays [r2 = 0.877, slope and intercept (+/- SE) = 0.88(+/- 0.051) and 0.316(+/- 0.523), respectively]. The range and mean (+/- SD) for the sandwich ELISA and the competitive EIA were 1.7-18.1 micrograms/L [8.7(+/- 4.4) micrograms/L] and 1.9-22.8 micrograms/L [9.1(+/- 4.4) micrograms/L], respectively.

Homogeneous enzyme immunoassay for cortisol with a centrifugal analyzer.

1984 ◽  
Vol 30 (11) ◽  
pp. 1824-1826 ◽  
Author(s):  
J M Izquierdo ◽  
A Quirós ◽  
J Alvarez-Uría ◽  
P Sotorrío
Keyword(s):  
Enzyme Immunoassay ◽  
Protein Complex ◽  
Acidic Solution ◽  
Serum Cortisol ◽  
Standard Curve ◽  
Analytical Recovery ◽  
Target Values ◽  
Assay Precision ◽  
Homogeneous Enzyme Immunoassay ◽  
Control Samples

Abstract We determine serum cortisol by a homogeneous enzyme immunoassay in the Cobas Bio centrifugal analyzer. To unbind cortisol from its protein complex, serum is treated for 15 min with an acidic solution. The reaction then proceeds automatically in the analyzer at 37 degrees C. To 50 microL of sample mixture is added 125 microL of reagent (cortisol antibodies, glucose 6-phosphate, and NAD+). This mixture is incubated for 60 s, after which 25 microL of a cortisol derivative labeled with glucose 6-phosphate is added; the increase in absorbance is monitored at 340 nm. The standard curve was linear from 10 to 500 micrograms of cortisol per liter. Within-assay precision (CV) varied from 0.2 to 0.6%, between-assay precision from 6.2 to 10.6%. Analytical recovery ranged from 100 to 103%. Results for control samples deviated from target values by 1.4 to 7.8%. Results compared well with those by radioimmunoassay. The method is reliable and practicable and will usefully replace previous routine methods for serum cortisol.

Indirect Competitive Enzyme Immunoassay for Tetrodotoxin Using a Biotin-Avidin System

1992 ◽  
Vol 75 (5) ◽  
pp. 883-886 ◽  
Author(s):  
Kendo Matsumura ◽  
Sadako Fukiya
Keyword(s):  
Bovine Serum Albumin ◽  
Enzyme Immunoassay ◽  
Immunoglobulin G ◽  
Bovine Serum ◽  
Keyhole Limpet Hemocyanin ◽  
Microtiter Plate ◽  
Mouse Bioassay ◽  
Detection Level ◽  
Standard Curve ◽  
Peroxidase Conjugate

Abstract An indirect competitive enzyme immunoassay was developed to determine tetrodotoxin (TTX). Antiserum against TTX was demonstrated in rabbits immunized with TTX-keyhole limpet hemocyanin conjugate. In this assay, TTX-bovine serum albumin was coated on the microtiter plate, which was incubated with standard TTX and biotinylated immunoglobulin G from antiserum. The amount of antibody bound on the surface of wells was determined by the reaction of avidin-peroxidase conjugate with its substrate. The minimal detection level of TTX ranged from 5 to 25 pg/assay. The standard curve was linear between 0.25 and 50 ng/assay. A good correlation (r= 0.974) for TTX was also obtained between this indirect competitive enzyme immunoassay and the mouse bioassay.

An enzyme immunoassay for galactosyltransferase isoenzyme II, and its clinical application to cancer diagnosis

1990 ◽  
Vol 36 (4) ◽  
pp. 598-601 ◽  
Author(s):  
M Uemura ◽  
M Yamasaki ◽  
S Yoshida ◽  
T Uezima ◽  
T Sakaguchi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Esophageal Cancer ◽  
Enzyme Immunoassay ◽  
Cancer Diagnosis ◽  
Blood Type ◽  
Benign Diseases ◽  
Standard Curve ◽  
Color Development ◽  
Sandwich Type ◽  
Normal Controls

Abstract In this "sandwich"-type enzyme immunoassay of galactosyltransferase isoenzyme II (GT-II), we used the monoclonal antibody MAb 3872 previously established and characterized to be specific for GT-II. Plastic beads coated with MAb 3872 were incubated with serum and buffer, washed, then incubated with MAb 3872 conjugated with horseradish peroxidase and again washed. Peroxidase activity remaining on the beads was measured by color development with o-phenylenediamine. The assay standard curve was linear from 0 to 150 kU/L. Inter- and intra-assay CVs were less than 9% and less than 7%, respectively. Mean GT-II values for 370 normal controls were 6.8 (SD 3.4) kU/L with no significant sex-, smoking-, or blood-type-related differences. We also assayed serum from patients with 11 different cancers or with matched benign diseases. The diagnostic sensitivity of GT-II for liver and esophageal cancer was 0.91 and 0.72, respectively, apparently higher than for carcinoembryonic antigen (CEA) or alpha-fetoprotein. GT-II and CEA concentrations in sera were not correlated. Evidently this assay may be useful as a cancer diagnostic test to supplement existing serological methods.

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