Comparison of a solid-phase, no-extraction radioimmunoassay for progesterone with an extraction assay for monitoring luteal function in the mare, bitch, and cow

ABSTRACT A rapid, solid-phase microtitre plate enzyme immunoassay (EIA) for progesterone is described using progesterone 3-O-carboxymethyloxime–horseradish peroxidase as the label and an antiserum raised in rabbits to a progesterone 11α-hemisuccinyl–bovine serum albumin immunogen. A competitive reaction was used with a reaction time of 2 h. Antibody-bound and free steroid were separated in a simple washing step of the antibody-adsorbed well surface. 2,2′-Azino-di-(3-ethylbenzthiazoline sulphonic acid) diammonium salt was used as the substrate with a reaction time of 1 h. A lower limit of sensitivity of 0·25 pg/well was obtained with the response being linear (logit/log) through 1000 pg/well. Results obtained by EIA and radioimmunoassay in several species gave excellent agreement (r = 0·98). This assay system allows accurate determination of progesterone in plasma with good specificity, precision and accuracy, and is suitable for the rapid assessment of luteal function and reproductive status in both clinical and research situations in a wide variety of species. J. Endocr. (1984) 101, 41–49

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Evaluation of a direct solid-phase radioimmunoassay for progesterone, useful for monitoring luteal function.

Clinical Chemistry ◽

10.1093/clinchem/30.2.284 ◽

1984 ◽

Vol 30 (2) ◽

pp. 284-286 ◽

Cited By ~ 49

Author(s):

N P Kubasik ◽

G D Hallauer ◽

R G Brodows

Keyword(s):

Detection Limit ◽

Luteal Phase ◽

Solid Phase ◽

Cross Reactivity ◽

Luteal Function ◽

Menstrual Cycles ◽

Analytical Recovery ◽

Solid Phase Radioimmunoassay ◽

Clinically Significant ◽

Direct Solid

Abstract We have evaluated a commercially available, direct, solid-phase radioimmunoassay kit for progesterone determination in serum or plasma. The assay is precise, within-run precision (CV) in the clinically significant ranges being 2.5 to 5.2%, between-run 5.5 to 5.8%. Mean analytical recovery of different concentrations of progesterone added to serum was 99.7% (range 95.3 to 102.7%). Fourteen closely related steroids showed no cross reactivity. The minimum detection limit was 0.5 microgram/L. Luteal-phase progesterone concentrations in serum were increased (greater than 3 micrograms/L) in 19 normal ovulatory menstrual cycles and decreased (less than 1.5 micrograms/L) in two nonovulatory cycles. We found this direct assay for progesterone to be analytically and clinically sound, and useful for assessing luteal-phase function.

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Determination of free progesterone in an ultrafiltrate of saliva collected in situ

Clinical Chemistry ◽

10.1093/clinchem/36.8.1488 ◽

1990 ◽

Vol 36 (8) ◽

pp. 1488-1493 ◽

Cited By ~ 11

Author(s):

W Schramm ◽

R H Smith ◽

P A Craig ◽

S H Paek ◽

H H Kuo

Keyword(s):

Solid Phase ◽

Biological Fluid ◽

Follicular Phase ◽

The Self ◽

Blood Samples ◽

Luteal Function ◽

Solid Phase Immunoassay ◽

Whole Saliva

Abstract We have investigated the utility of an ultrafiltrate of saliva for measuring progesterone as an indicator of luteal function during the menstrual cycle of women. A filtrate of saliva is collected in the mouth by means of an osmotic pump that accumulates medium containing only molecules less than 12,000 Da. We analyzed the nonextracted ultrafiltrate by a solid-phase immunoassay for progesterone and monitored the mid-luteal surge of lutropin in urine with a liquid-phase radioimmunoassay. Progesterone concentrations in the ultrafiltrate are significantly lower during the follicular phase and increase after the release of lutropin. The concentration of progesterone in the ultrafiltrate correlates closely with total progesterone in matched blood samples (r = 0.84, cycle 1; and r = 0.89, cycle 2). Likewise, we found a good correlation between the results in whole saliva and in the ultrafiltrate (r = 0.95). The described method of obtaining a pre-processed specimen noninvasively simplifies the self-collection of samples by patients (including collection at home); excludes potential interference from microorganisms, desquamated cells, and salivary components; and simplifies the processing of the biological fluid in the laboratory.

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Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111240 ◽

1984 ◽

Vol 42 ◽

pp. 226-227 ◽

Cited By ~ 1

Author(s):

K. Pegg-Feige ◽

F. W. Doane

Keyword(s):

Electron Microscopy ◽

Cell Culture ◽

Immunoelectron Microscopy ◽

Detection Method ◽

Solid Phase ◽

Protein A ◽

Plant Viruses ◽

Rapid Diagnosis ◽

Virus Diagnosis ◽

Antiviral Antibody

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.

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Application of solid-phase immune electron microscopy to study physical and stability characteristics of enterically transmitted non-a, non-b hepatitis virus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100102481 ◽

1988 ◽

Vol 46 ◽

pp. 80-81

Author(s):

Charles D. Humphrey ◽

E. H. Cook ◽

Karen A. McCaustland ◽

Daniel W. Bradley

Keyword(s):

Electron Microscopy ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Solid Phase ◽

Etiologic Agent ◽

World Health ◽

Health Concern ◽

Morphological Identification ◽

Immune Electron Microscopy

Enterically transmitted non-A, non-B hepatitis (ET-NANBH) is a type of hepatitis which is increasingly becoming a significant world health concern. As with hepatitis A virus (HAV), spread is by the fecal-oral mode of transmission. Until recently, the etiologic agent had not been isolated and identified. We have succeeded in the isolation and preliminary characterization of this virus and demonstrating that this agent can cause hepatic disease and seroconversion in experimental primates. Our characterization of this virus was facilitated by immune (IEM) and solid phase immune electron microscopic (SPIEM) methodologies.Many immune electron microscopy methodologies have been used for morphological identification and characterization of viruses. We have previously reported a highly effective solid phase immune electron microscopy procedure which facilitated identification of hepatitis A virus (HAV) in crude cell culture extracts. More recently we have reported utilization of the method for identification of an etiologic agent responsible for (ET-NANBH).

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Identification of hepatitis a virus in cell cultures by solid phase immune electron microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100142815 ◽

1986 ◽

Vol 44 ◽

pp. 238-239

Author(s):

C.D. Humphrey ◽

T.L. Cromeans ◽

E.H. Cook ◽

D.W. Bradley

Keyword(s):

Electron Microscopy ◽

Liquid Phase ◽

Cell Cultures ◽

Rapid Detection ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Solid Phase ◽

Immune Electron Microscopy ◽

Detection And Identification

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.

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Dynamic studies of silicide-mediated crystallization of amorphous silicon

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100131395 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1352-1353

Author(s):

C. Hayzelden ◽

J. L. Batstone

Keyword(s):

Ion Implantation ◽

Solid Phase ◽

Metallic Phase ◽

Single Crystal Substrate ◽

Free Electrons ◽

Melt Temperature ◽

Transmission Electron ◽

Crystal Substrate ◽

High Resolution Tem

Epitaxial reordering of amorphous Si(a-Si) on an underlying single-crystal substrate occurs well below the melt temperature by the process of solid phase epitaxial growth (SPEG). Growth of crystalline Si(c-Si) is known to be enhanced by the presence of small amounts of a metallic phase, presumably due to an interaction of the free electrons of the metal with the covalent Si bonds near the growing interface. Ion implantation of Ni was shown to lower the crystallization temperature of an a-Si thin film by approximately 200°C. Using in situ transmission electron microscopy (TEM), precipitates of NiSi2 formed within the a-Si film during annealing, were observed to migrate, leaving a trail of epitaxial c-Si. High resolution TEM revealed an epitaxial NiSi2/Si(l11) interface which was Type A. We discuss here the enhanced nucleation of c-Si and subsequent silicide-mediated SPEG of Ni-implanted a-Si.Thin films of a-Si, 950 Å thick, were deposited onto Si(100) wafers capped with 1000Å of a-SiO2. Ion implantation produced sharply peaked Ni concentrations of 4×l020 and 2×l021 ions cm−3, in the center of the films.

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TEM characterization of Au/Polysilicon interactions

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100176381 ◽

1990 ◽

Vol 48 (4) ◽

pp. 650-651

Author(s):

N. David Theodore ◽

Leslie H. Allen ◽

C. Barry Carter ◽

James W. Mayer

Keyword(s):

Solid Phase ◽

Single Crystal Silicon ◽

Chemical Vapor ◽

Electrical Contacts ◽

Silicon Substrates ◽

Crystal Silicon ◽

Transmission Electron ◽

Polysilicon Films ◽

The Stability

Metal/polysilicon investigations contribute to an understanding of issues relevant to the stability of electrical contacts in semiconductor devices. These investigations also contribute to an understanding of Si lateral solid-phase epitactic growth. Metals such as Au, Al and Ag form eutectics with Si. reactions in these metal/polysilicon systems lead to the formation of large-grain silicon. Of these systems, the Al/polysilicon system has been most extensively studied. In this study, the behavior upon thermal annealing of Au/polysilicon bilayers is investigated using cross-section transmission electron microscopy (XTEM). The unique feature of this system is that silicon grain-growth occurs at particularly low temperatures ∽300°C).Gold/polysilicon bilayers were fabricated on thermally oxidized single-crystal silicon substrates. Lowpressure chemical vapor deposition (LPCVD) at 620°C was used to obtain 100 to 400 nm polysilicon films. The surface of the polysilicon was cleaned with a buffered hydrofluoric acid solution. Gold was then thermally evaporated onto the samples.

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