GB2051357A - Simultaneous Detection of Indicators of Hepatitis Virus Exposure - Google Patents
Simultaneous Detection of Indicators of Hepatitis Virus Exposure Download PDFInfo
- Publication number
- GB2051357A GB2051357A GB8008095A GB8008095A GB2051357A GB 2051357 A GB2051357 A GB 2051357A GB 8008095 A GB8008095 A GB 8008095A GB 8008095 A GB8008095 A GB 8008095A GB 2051357 A GB2051357 A GB 2051357A
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- hepatitis
- reagent
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- markers
- antibody
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
- G01N33/5761—Hepatitis B
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- Communicable Diseases (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
A method for simultaneously detecting in a sample at least two different markers (a marker is an antigen or antibody) evidencing exposure to hepatitis virus comprises contacting the sample with a solid phase reagent which is coated with at least two different, non- complementary immunoreactants which are complementary to the unknown markers to be detected, then with a liquid reagent comprising at least two different hepatitis markers or immunoreactants, each selected to either react or compete with one of the unknown markers and each labelled with a detectably distinct tag.
Description
SPECIFICATION
Method and Reagents for Simultaneously
Detecting Different Markers of Hepatitis
This invention discioses an improvement in solid phase immunoassay methods for the detection and determination of antigens and antibodies (markers) relating to hepatitis.
There are at least two distinct types of viral hepatitis. Hepatitis A is believed to be caused by the hepatitis A antigen (HAVAg) and is generally characterized by an incubation period of two to six weeks, mild prodromata and a relatively mild clinical illness. The disease is generally transmitted by contaminated food or liquid, but has also been shown to be transmitted by systemic inoculation. Hepatitis A is frequently called "infectious hepatitis", and in the United
States the number of reported cases is over 50,000 annually.
The hepatitis B virus is believed to be the most probable etiologic agent for "serum hepatitis".
Hepatitis B infection is generally transmitted by blood products or contaminated instruments such as needles, but it may also be transmitted by other means. Previously, a hepatitis B infection was associated with an incubation period ranging from six weeks to six months. However, incubation periods as short as two weeks have been documented. The illness may be mild or asymptomatic, but if symptomatic, manifestations may be especially severe. Prodromata may include arthralgias, arthritis, rash, fever, anorexia, fatigue and pruritis with or without jaundice.
At least three distinct antigen-antibody systems can be associated with hepatitis virus B: the surface (HBsAg: anti-HBs), the "core" (HBcAg: anti-HBc), and e-antigen (HBeag: anti-HBe). The hepatitis B surface antigen (HBsAg) is found in the blood as 22 nm spheres and as elongated tubules which are 22 nm in diameter and variable in length and is believed to represent the protein coat of the hepatitis B virus.
A 42 nm particle containing DNA and a DNA polymerase is considered to represent the infectious virus (Dane particle). The surface of the
Dane particle is similar or identical to HBsAg. In detergents, the Dane particle is degraded to a 27 nm electron dense core, HBCAg. The latter is seen in nuclei of hepatocytes of patients with serum hepatitis during the acute infection stage.
Thus, patients with viral hepatitis type B might be expected to produce antibodies to the surface antigen (anti-H B6), and also to the protein core (anti-HBc). HBsAg in serum has been a consistent marker for ths presence of the hepatitis B virus, and anti-HBc usually appears during early viremia, often accompanying antigenemia (HB6Ag), at the height of liver disfunction and long before the appearance of anti-HB . Anti- HBC is generally associated with prolonged circulation of HBsAg suggesting that anti-HBc is produced in response to the active replication of the virus.
In 1972, the hepatitis e antigen (HBeAg) was detected and characterized. The e marker has been found in HBsAg-positive serum and is thought to occur more commonly in serum of chronic HBsAg carriers with active liver disease than in healthy carriers. In patients, during the incubation period of hepatitis B, the e antigen was shown to appear just after the appearance of HBsAg and before clinically apparent liver injury.
Logically, its presence in such sera would be indicative of a poor prognosis and on-going liver damage. Conversely, the presence of e antibody (anti-H Be) would be indicative of a good prognosis. These correlations are not absolute, but may be useful clinical guides.
Since the serologic markers for hepatitis appear in consistent, sequential order during the course of infection, acute disease and recovery, an analysis for two or more markers would be valuable to assess the time-course of the disease.
An assay for more than one marker can also provide an important double-check for HB5Ag in blood doners or patients in an effort to reduce the incidence of type-B hepatitis. Such a need has been demonstrated by Goldfield et al; Am. J. Med.
Sci., 270: 335-342 (1975), who in careful follow-ups of recipients of blood negative for HBsAg, found evidence of exposure to the hepatitis antigen in 7 of 465 patients. Clearly, a method of detecting more than one marker to the hepatitis B virus should decrease or prevent the occurrence of false negatives. Similarly, in many instances a positive response to two tests minimizes the possibility of a false positive occurring in one test.
While the reagents and methods described and claimed herein are similar to known commercial products and procedures for detecting the various markers to viral hepatitis, heretofore there has been no disclosure or suggestion describing any techniques for detecting said markers simultaneously.
Accordingly, this specification describes in detail reagents and a method useful for detecting simultaneously in a sample at least two different markers evidencing exposure to hepatitis virus.
The method comprises contacting the sample with a solid support which has been coated with at least two different, non-complimentary immunoreactants which are complimentary to the unknown markers; incubating the sample with the coated solid support for a period up to 24 hours;
separating the coated solid support from the sample;
washing the coated solid support;
contacting the washed solid support with a liquid reagent comprising at least two different hepatitis markers or immunoreactants, each selected to either react or compete with one of the unknown hepatitis markers and each labeled with detectably distinct tags;
separating the solid phase from the liquid reagent; and
determining the presence of labeled markers on the solid support by detecting each distinct tag.
The following examples will demonstrate the preparation and use of representative reagents within the scope of the claimed invention. The first example describes the method of preparing a solid support coated with two different noncomplimentary immunoreactants which are complimentary to markers evidencing exposure to hepatitis.
More specifically, a solid support (bead) has been coated with two different, noncomplimentary immunoreactants in such a manner that both retain their reactivity and are able to combine with complimentary markers in an unknown specimen.
In the first example a polystyrene bead has been coated with an antibody to HBsAg and with the core antigen, HBCAg. The antibody retains its avidity and will react with any HBsAg in the unknown specimen and the affixed core antigen will retain its antigenicity and react with any antibody to the core (anti-HBc) in the unknown specimen. The crux of this invention is that both immunoreactions will occur simultaneously with a singie solid phase reagent.
Example I
Preparation of Coated Support
A formulation of HBCAg was made by exposing
Dane particles to a solution of 2% 2mercaptoethanol (2-ME) and 5% Tween 80 in
Tris-EDTA-saline (TSE) buffers at 37 OC for 2 hours. That solution was diluted lO-fold in 5% 2ME-TSE and allowed to stand overnight before diluting to final concentration of 1:8000 in TSE.
Separately, polystyrene beads (1/4") were coated with a 1:2000 dilution of anti-HBs serum (guinea pig) in phosphate buffered saline (PBS) by soaking for two hours. The beads were removed, washed and dried. The core preparation was then poured over the coated beads and the core antigen was allowed to adhere to the bead for 2 days at room temperature. The beads were removed from the buffer solution, washed, coated with a 10% sucrose solution to stabilize the adherents and air dried.
Although polystyrene beads are preferred because they are easily coated and manipulated, any solid support of either macro or micro dimensions, fashioned from a variety of plastics, metal and glass could just as easily be coated and used to demonstrate the claimed invention.
While it is possible that the beads may be coated first with core antigen and subsequently coated with anti-HBs, it has been observed that coating the beads first with the guinea pig anti HB, serum significantly augments the adherence of core antigen.
It is also important to note that other combinations of non-complimentary immunoreactants relating to hepatitis virus may also be employed. The immunoreactants must, of course, be non-complimentary because a complimentary attraction would diminish the antigenicity and avidity of the reagent.
The following example will demonstrate the preparation of a liquid phase reagent containing
an immunoreactant, anti-HBs, tagged with a radio
label (1251) and a marker, anti-HBc, tagged with a
reactive enzyme label (horseradish peroxidase).
While the terms "marker" and
"immunoreactant" might be used interchangably, it is preferable if the term "marker" is used to designate the antigen or antibody, and their
equivalents, to be detected in the unknown sample. The term "immunoreactant" is used to
define either the antigen or antibody complimentary to the markers to be detected.
Accordingly, in the following examples, the antigen and antibody employed on the solid support are always complementary to one of the
unknown markers, and, therefore, are referred ta as immunoreactants.
In the liquid phase reagent, the labeled surface antibody is complimentary to the unknown marker (HBsAg) and is, therefore, referred to as the labeled immunoreactant. The labeled core antibody, however, is identical to the marker to be detected and is, therefore, referred to as a labeled marker.
Example II
Tagged Antibody Reagent
The iodination (1251) of antibody to HBsAg (anti HB9) entailed adding approximately 0.1 ml of a 0.5 M phosphate buffer at pH7.5, a small volume of 5-6 mci of Na1251, and 100 mg. of purified anti-HBs to a one dram vial, adjusting the pH to 7.5-8.0 and mixing all ingredients. To this mixture was added 50 microliters of a freshly prepared solution of chloramine T (35 mg in 10 ml. 0.05 M phosphate buffer, pH 7.5). After additional mixing, the reaction was allowed to proceed at room temperature for 60 seconds.
Fifty microliters of a freshly prepared solution of sodium metalsulfite (35 mg in 10 ml of 0.05 M phosphate buffer at pH 7.5) were added to the reaction to reduce the chloramine T and thereby stop the reaction.
The reaction efficiency was checked by placing 5 microliters of the reaction mixture on a strip of
Whatman No. 1 paper strip and chromatographing the mixture in 70% methanol.
The iodinated protein remained at the origin while the free 1251 migrated with the solvent. The % of 1251 in protein is considered to be the % of reaction efficiency.
The crude iodinated antibody was purified using a Sephadex G-50 gel column with 0.1 molar Tris buffer containing 0.9% sodium chloride at pH 7.8. The column was pretreated with a small volume of a 30% aqueous solution of bovine serum albumin followed by an equal additional volume of 0.1 molar Tris buffer solution containing 0.9% sodium chloride at pH 7.8. The iodinated antibody was added to the top of the column and washed through using the saline buffered Tris solution. The labeled antibody was the first eluate collected from the column.
The conjugation of antibody to hepatitis B core to horseradish peroxidase (HRP) entailed activating peroxidase by using sodium meta periodate. The excess periodate and by-products
were separated from the active peroxidase by gel
filtration (sephadex G-25) column. The activated
peroxidase was then reacted with the antibody (anti-HBc). The reaction occurred spontaneously.
Sodium borohydride was then added to stabilize
the bond formed between peroxidase and the
antibody, and acetone was added to destroy the
remaining sodium borohydride.
When employed according to the teaching of
this invention, the two tagged antibodies are
diluted into a diluent containing:
50% fetal calf serum 15% normal human serum
.005 M EDTA 0.1% Tween-20
0.01%Thiomersal in PBS
It should be apparent that a variety of
detectable tags may be employed. The only
requirement, naturally, is that they be detectably
distinct. Any of a variety of isotopes could be
utilized just as easily as '251. It is not necessary
that one marker or immunoreactant be radio
labeled and the other enzymatically orflorescently labeled since different isotopes are, themselves,
detectably distinct.Similarly, all labeled
components of the reagent might just as easily
employ different enzymes requiring different
substrates yielding detectably distinct reaction
products.
It should be noted that any of the antibodies
and antigens of hepatitis A and B could be labeled
to perform according to the disclosed invention.
The only immunochemical requirements are that
the labeled components not be complimentary
with each other or to the markers to be
determined.
Example Ill
Simultaneous Detection of HBsAg and Anti
HBc
Serum samples containing the "unknown"
hepatitis markers were added to the individual
wells of a reaction tray and a polystyrene bead
coated in accordance with Example I was added
to each specimen sample and allowed to incubate
for 2 hours at 450C. The beads were removed
from the reaction well, washed with water to
remove any excess reagent, and added to 0.2 ml
of the liquid phase reagent prepared in
accordance with Example II. The solid and liquid
phases were incubated for a period of one hour at
450C. The solid phase was then separated from
the antibody reagent and washed four times with
water to remove excess reagent.The washed
beads were placed in test tubes containing o- phenylenediamine (30 mg in 10 ml of 0.1 M
citrate buffer, pH 5.5) and allowed to incubate for
30 minutes. Following that period the enzyme
reaction was stopped by the addition of 1 ml of 1
M HCI and the tubes were visually examined for
the presence or absence of color resulting from
the reaction of the enzyme tag and the substrate.
The absence or low incidence of color indicated
that anti-HBc was present in the unknown sample and competed with the labeled marker for reaction sites on the coated support. Next, radioactivity on the beads was counted in a gamma counter and the CPM's was recorded.
Radioactivity on the bead indicated that there was HBsAg in the unknown sample which adhered to the affixed antibody and provided a binding site for the radio-labeled antibody.
Claims (13)
1. A method of simultaneously detecting in a sample at least two different markers evidencing exposure to hepatitis virus, said method comprising:
a) contacting the sample with a solid phase coated with at least two different, noncomplimentary immunoreactants complimentary to the unknown markers;
b) incubating the sample with the coated solid phase for a period of 1-24 hours;
c) separating the coated solid phase from the sample;
d) washing the coated solid phase to remove unbound sample;
e) contacting the washed solid support with a liquid reagent comprising at least two different hepatitis markers or immunoreactants, each selected to either react or compete with one of the known hepatitis markers, and each labeled with detectably distinct tags;
f) separating the solid phase from the labeled reagent;;
g) and determining the presence of labeled markers or immunoreactants on the solid support by detecting the distinct tags.
2. A method of simultaneously detecting in a sample hepatitis B surface antigen and antibody to hepatitis B core antigen, said method comprising:
a) contacting the sample with a solid phase coated with both hepatitis B core antigen and an antibody to hepatitis B surface antigen;
b) incubating the sample with the coated solid phase for a period of 1-24 hours;
c) separating the coated solid phase from the sample;
d) washing the coated solid phase to remove unbound sample;
e) contacting the washed solid support with a liquid reagent comprising antibodies to hepatitis B surface antigen and hepatitis B core antigen each labeled with detectably distinct tags.
f) separating the solid phase from the labeled antibody reagent; and
g) determining the presence of labeled antibodies on the solid support by detecting the distinct tags.
3. The method according to Claim 2 wherein the antibody to the surface antigen is radiolabeled and the antibody to the core antigen is labeled with an enzyme.
4. The method according to Claim 3 wherein the radiolabel is 1251 and the enzyme is horseradish peroxidase.
5. A reagent useful for simultaneously detecting in a sample at least two different markers evidencing exposure to hepatitis virus said reagent comprising:
a solid support coated with at least two different non-complimentary immunoreactants complimentary to the unknown markers.
6. A reagent according to Claim 5 wherein the solid support is further defined as being coated with hepatitis B core antigen and an antibody to hepatitis B surface antigen.
7. A reagent useful for simultaneously detecting in a sample at least two different markers evidencing exposure to hepatitis virus said reagent comprising:
at least two different hepatitis markers or immunoreactants each selected to either react or compete with an unknown hepatitis marker and each labeled with detectably distinct tags.
8. A reagent according to Claim 7 wherein the reagent is further defined as comprising antibodies to hepatitis B surface antigen and hepatitis B core antigen.
9. A reagent according to Claim 8 wherein the antibody to hepatitis B surface antigen is radiolabeled.
10. A reagent according to Claim 9 wherein the radiolabel is 1251..
11. A reagent according to Claim 8 wherein the antibody to hepatitis B core antigen is labeled with an enzyme.
12. A reagent according to Claim 11 wherein the enzyme label is horseradish peroxidase.
13. A reagent according to claim 5, prepared according to any one of the Examples herein.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US4831979A | 1979-06-14 | 1979-06-14 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| GB2051357A true GB2051357A (en) | 1981-01-14 |
| GB2051357B GB2051357B (en) | 1983-05-18 |
Family
ID=21953924
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| GB8008095A Expired GB2051357B (en) | 1979-06-14 | 1980-03-11 | Simultaneous detection of sindicators of hepatitis virus exposure |
Country Status (12)
| Country | Link |
|---|---|
| JP (2) | JPS562558A (en) |
| AT (1) | AT368813B (en) |
| AU (1) | AU530580B2 (en) |
| BE (1) | BE883824A (en) |
| CA (1) | CA1148859A (en) |
| CH (1) | CH654113A5 (en) |
| DE (1) | DE3021891C2 (en) |
| FR (1) | FR2459482A1 (en) |
| GB (1) | GB2051357B (en) |
| IT (1) | IT1140846B (en) |
| NL (1) | NL8001661A (en) |
| ZA (1) | ZA801318B (en) |
Cited By (20)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0058428A2 (en) * | 1981-02-18 | 1982-08-25 | Eisai Co., Ltd. | An enzyme immuno-assay for simultaneously measuring a plurality of samples and test vessel for carrying out this method |
| GB2125547A (en) * | 1982-07-31 | 1984-03-07 | Mochida Pharm Co Ltd | Simultaneous immunoassay of two or more substances |
| EP0108992A1 (en) * | 1982-11-05 | 1984-05-23 | Kabushiki Kaisha Toshiba | Immunoassay |
| EP0131974A1 (en) * | 1983-06-08 | 1985-01-23 | Akzo N.V. | Determination of delta-antigens in body fluids |
| US4508830A (en) * | 1982-02-10 | 1985-04-02 | Baker Terence S | Assay |
| GB2165046A (en) * | 1982-02-10 | 1986-04-03 | Baker Terence S | Ligand molecule |
| US4701410A (en) * | 1983-09-14 | 1987-10-20 | Akzo N.V. | Method for the immunochemical determination of hepatitis B core antigens |
| US4701421A (en) * | 1984-08-27 | 1987-10-20 | Akzo, N.V. | Determination of protecting anti-HBV immunoglobulins |
| EP0286264A2 (en) * | 1987-03-23 | 1988-10-12 | Ortho Pharmaceutical Corporation | Assay for detecting hepatitis antigens and aids antibodies |
| WO1990004182A1 (en) * | 1988-10-14 | 1990-04-19 | Virovahl S.A. | A method for simultaneous detection of different types of antibodies and/or antigens produced by individual cells |
| EP0351248A3 (en) * | 1988-07-14 | 1990-11-22 | Idexx Corp. | Detection of an antibody and antigen in an immunoassay |
| EP0402757A2 (en) * | 1989-06-16 | 1990-12-19 | Biochrom Beteiligungs GmbH & Co. Produktionsgesellschaft | Solid phase immunoassay and kit for the determination of antigens and antibodies |
| EP0408542A2 (en) * | 1989-07-13 | 1991-01-16 | IMMUNO Aktiengesellschaft | A method of determining antigens and/or antibodies in human body liquids, and a test kit for carrying out this method |
| EP0473065A2 (en) * | 1990-08-29 | 1992-03-04 | Abbott Laboratories | Simultaneous assay for detecting two or more analytes |
| EP1310512A2 (en) * | 2001-11-11 | 2003-05-14 | Ortho-Clinical Diagnostics, Inc. | Hcv core protein sequences |
| US6855809B2 (en) * | 2001-06-26 | 2005-02-15 | Abbott Laboratories | Methods for the simultaneous detection of HCV antigens and HCV antibodies |
| US7101683B2 (en) | 2001-06-26 | 2006-09-05 | Abbott Laboratories | Methods for the simultaneous detection of HCV antigens and HCV antibodies |
| EP1672366A3 (en) * | 1999-06-11 | 2007-11-28 | THE GOVERNMENT OF THE UNITED STATES OF AMERICA, represented by THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES | Methods and compositions for opsonophagocytic assays |
| US7316905B1 (en) * | 1998-07-30 | 2008-01-08 | Advanced Life Science Institute, Inc. | Method for measurement of hepatitis C virus |
| WO2014176535A1 (en) | 2013-04-26 | 2014-10-30 | Bio-Rad Laboratories, Inc. | Multiplex hepatitis b assay |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS58187861A (en) * | 1982-04-26 | 1983-11-02 | Otsuka Pharmaceut Co Ltd | Assay of antibody related to adult type t leukemia |
| GB8607101D0 (en) * | 1986-03-21 | 1986-04-30 | Serono Diagnostics Ltd | Immunoassay |
| DE4236189A1 (en) * | 1992-10-27 | 1994-04-28 | Boehringer Mannheim Gmbh | Method for the simultaneous determination of antigens and antibodies |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3867517A (en) * | 1971-12-21 | 1975-02-18 | Abbott Lab | Direct radioimmunoassay for antigens and their antibodies |
| GB1506017A (en) * | 1974-03-04 | 1978-04-05 | Int Diagnostic Tech | Fluorometric system and method for the detection of biologically derived substances |
| SE7610683L (en) * | 1975-09-29 | 1977-06-10 | Cordis Corp | METHOD OF DETERMINING THE NERVARON OF AN ANTIGEN ASSOCIATE WITH HEPATITIS |
| AT343822B (en) * | 1976-08-20 | 1978-06-26 | Immuno Ag | RADIOIMMUNOLOGICAL METHOD AND EQUIPMENT FOR DETERMINING ANTIGENES |
| DE2803154C2 (en) * | 1977-01-27 | 1986-07-24 | Becton, Dickinson and Co., Paramus, N.J. | Simultaneous radio testing of folate and vitamin B 12 |
| CA1100037A (en) * | 1977-03-11 | 1981-04-28 | Chung-Mei Ling | Hb.sub.c ag coated on solid phase |
| US4102996A (en) * | 1977-04-20 | 1978-07-25 | Merck & Co., Inc. | Method of preparing hepatitis B core antigen |
| US4189464A (en) * | 1978-05-05 | 1980-02-19 | Institute For Cancer Research | Hepatitis B testing reagent and method |
| IL55816A (en) * | 1978-10-30 | 1982-04-30 | Ames Yissum Ltd | Method for simultaneous immunoassay of several different antibodies and a kit therefor |
-
1980
- 1980-02-29 CA CA000346729A patent/CA1148859A/en not_active Expired
- 1980-03-06 ZA ZA00801318A patent/ZA801318B/en unknown
- 1980-03-06 AU AU56226/80A patent/AU530580B2/en not_active Ceased
- 1980-03-11 GB GB8008095A patent/GB2051357B/en not_active Expired
- 1980-03-20 NL NL8001661A patent/NL8001661A/en not_active Application Discontinuation
- 1980-05-07 IT IT21874/80A patent/IT1140846B/en active
- 1980-06-06 JP JP7568180A patent/JPS562558A/en active Granted
- 1980-06-11 DE DE3021891A patent/DE3021891C2/en not_active Expired
- 1980-06-12 AT AT0309980A patent/AT368813B/en not_active IP Right Cessation
- 1980-06-13 FR FR8013217A patent/FR2459482A1/en active Granted
- 1980-06-13 CH CH4579/80A patent/CH654113A5/en not_active IP Right Cessation
- 1980-06-13 BE BE0/201040A patent/BE883824A/en not_active IP Right Cessation
-
1990
- 1990-01-16 JP JP2004676A patent/JPH02276969A/en active Pending
Cited By (31)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0058428A3 (en) * | 1981-02-18 | 1983-01-12 | Eisai Co., Ltd. | Method for measuring substances by enzyme immunoassay and test vessel for carrying out this method |
| EP0058428A2 (en) * | 1981-02-18 | 1982-08-25 | Eisai Co., Ltd. | An enzyme immuno-assay for simultaneously measuring a plurality of samples and test vessel for carrying out this method |
| US4508830A (en) * | 1982-02-10 | 1985-04-02 | Baker Terence S | Assay |
| GB2165046A (en) * | 1982-02-10 | 1986-04-03 | Baker Terence S | Ligand molecule |
| GB2125547A (en) * | 1982-07-31 | 1984-03-07 | Mochida Pharm Co Ltd | Simultaneous immunoassay of two or more substances |
| EP0108992A1 (en) * | 1982-11-05 | 1984-05-23 | Kabushiki Kaisha Toshiba | Immunoassay |
| US4623618A (en) * | 1982-11-05 | 1986-11-18 | Tokyo Shibaura Denki Kabushiki Kaisha | Simultaneous quantitative immunoassay for different antigens or antibodies |
| EP0131974A1 (en) * | 1983-06-08 | 1985-01-23 | Akzo N.V. | Determination of delta-antigens in body fluids |
| US4701410A (en) * | 1983-09-14 | 1987-10-20 | Akzo N.V. | Method for the immunochemical determination of hepatitis B core antigens |
| US4701421A (en) * | 1984-08-27 | 1987-10-20 | Akzo, N.V. | Determination of protecting anti-HBV immunoglobulins |
| EP0286264A2 (en) * | 1987-03-23 | 1988-10-12 | Ortho Pharmaceutical Corporation | Assay for detecting hepatitis antigens and aids antibodies |
| EP0286264A3 (en) * | 1987-03-23 | 1988-10-19 | Ortho Pharmaceutical Corporation | assay for detecting hepatitis antigens and aids antibodies |
| EP0351248A3 (en) * | 1988-07-14 | 1990-11-22 | Idexx Corp. | Detection of an antibody and antigen in an immunoassay |
| US5627026A (en) * | 1988-07-14 | 1997-05-06 | Idexx Laboratories, Inc. | Detection of both an antibody and an antigen in a single sample aliquot |
| WO1990004182A1 (en) * | 1988-10-14 | 1990-04-19 | Virovahl S.A. | A method for simultaneous detection of different types of antibodies and/or antigens produced by individual cells |
| EP0402757A2 (en) * | 1989-06-16 | 1990-12-19 | Biochrom Beteiligungs GmbH & Co. Produktionsgesellschaft | Solid phase immunoassay and kit for the determination of antigens and antibodies |
| EP0402757A3 (en) * | 1989-06-16 | 1991-10-30 | Biochrom Beteiligungs GmbH & Co. Produktionsgesellschaft | Solid phase immunoassay and kit for the determination of antigens and antibodies |
| EP0408542A3 (en) * | 1989-07-13 | 1993-01-13 | Immuno Aktiengesellschaft Fuer Chemisch-Medizinische Produkte | A method of determining antigens and/or antibodies in human body liquids, and a test kit for carrying out this method |
| EP0408542A2 (en) * | 1989-07-13 | 1991-01-16 | IMMUNO Aktiengesellschaft | A method of determining antigens and/or antibodies in human body liquids, and a test kit for carrying out this method |
| EP0473065A2 (en) * | 1990-08-29 | 1992-03-04 | Abbott Laboratories | Simultaneous assay for detecting two or more analytes |
| EP0473065A3 (en) * | 1990-08-29 | 1992-08-26 | Abbott Laboratories | Simultaneous assay for detecting two or more analytes |
| US7316905B1 (en) * | 1998-07-30 | 2008-01-08 | Advanced Life Science Institute, Inc. | Method for measurement of hepatitis C virus |
| EP1672366A3 (en) * | 1999-06-11 | 2007-11-28 | THE GOVERNMENT OF THE UNITED STATES OF AMERICA, represented by THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES | Methods and compositions for opsonophagocytic assays |
| US6855809B2 (en) * | 2001-06-26 | 2005-02-15 | Abbott Laboratories | Methods for the simultaneous detection of HCV antigens and HCV antibodies |
| US7101683B2 (en) | 2001-06-26 | 2006-09-05 | Abbott Laboratories | Methods for the simultaneous detection of HCV antigens and HCV antibodies |
| EP2397566A1 (en) * | 2001-06-26 | 2011-12-21 | Abbott Laboratories | Methods for the simultaneous detection of HCV antigens and HCV antibodies |
| EP1310512A3 (en) * | 2001-11-11 | 2004-01-21 | Ortho-Clinical Diagnostics, Inc. | Hcv core protein sequences |
| EP1310512A2 (en) * | 2001-11-11 | 2003-05-14 | Ortho-Clinical Diagnostics, Inc. | Hcv core protein sequences |
| US7332269B2 (en) | 2001-11-11 | 2008-02-19 | Ortho-Clinical Diagnostics, Inc. | HCV core protein sequences |
| WO2014176535A1 (en) | 2013-04-26 | 2014-10-30 | Bio-Rad Laboratories, Inc. | Multiplex hepatitis b assay |
| US9927439B2 (en) | 2013-04-26 | 2018-03-27 | Bio-Rad Laboratories, Inc. | Multiplex hepatitis B assay |
Also Published As
| Publication number | Publication date |
|---|---|
| FR2459482A1 (en) | 1981-01-09 |
| NL8001661A (en) | 1980-12-16 |
| ZA801318B (en) | 1981-02-25 |
| JPH0220945B2 (en) | 1990-05-11 |
| DE3021891C2 (en) | 1983-08-04 |
| AT368813B (en) | 1982-11-10 |
| IT8021874A0 (en) | 1980-05-07 |
| FR2459482B1 (en) | 1984-10-26 |
| JPH02276969A (en) | 1990-11-13 |
| JPS562558A (en) | 1981-01-12 |
| AU530580B2 (en) | 1983-07-21 |
| IT1140846B (en) | 1986-10-10 |
| ATA309980A (en) | 1982-03-15 |
| BE883824A (en) | 1980-12-15 |
| CH654113A5 (en) | 1986-01-31 |
| CA1148859A (en) | 1983-06-28 |
| AU5622680A (en) | 1980-12-18 |
| DE3021891A1 (en) | 1980-12-18 |
| GB2051357B (en) | 1983-05-18 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PCNP | Patent ceased through non-payment of renewal fee |
Effective date: 19950311 |