Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Reexamination Certificate
2000-10-12
2002-12-03
Stucker, Jeffrey (Department: 1648)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
C424S263100, C435S004000, C435S007200, C435S007320, C435S252300, C435S320100
Reexamination Certificate
active
06489122
ABSTRACT:
FIELD OF THE INVENTION
The invention relates to
Chlamydia pneumoniae
antigenic polypeptides, fused proteins containing the polypeptides, DNAs coding therefor, recombinant vectors carrying the DNAs, transformants containing the recombinant vectors, a method for production of antibody, a method and reagents for detection and/or measurement of antibody, a method and agents for diagnosis of
Chlamydia pneumoniae
infections, probes and primers for detection and/or measurement of
Chlamydia pneumoniae
gene, and a method and reagents for detection and/or measurement of
Chlamydia pneumoniae
gene. The invention can be effectively used in the pharmaceutical industry, particularly in the preparation of agents for diagnosis of
Chlamydia pneumoniae
infections.
BACKGROUND ART
Several kinds of species are known in Chlamydia, that is,
Chlamydia trachomatis, Chlamydia psittaci, Chlamydia pecorum, Chlamydia pneumoniae
and the like.
Chlamydia trachomatis
causes trachoma, venereal lymphogranuloma, urogenital infections, inclusion conjunctivitis, neonatal pneumonia and the like.
Chlamydia psittaci
causes psittocosis and the like.
Chlamydia pneumoniae
causes respiratory infections, atypical pneumonia and the like.
Since the symptoms of infections in the respiratory apparatus which are caused by
Chlamydia pneumoniae
are similar to those of infections caused by
Mycoplasma pneumoniae
or Influenza virus, physicians often make a wrong diagnosis. Hence, there is a need for the development of a simple method for diagnosing the infections caused by
Chlamydia pneumoniae.
In general, an infection can reliably be diagnosed by detecting the causative bacterium in the infected site or by detecting an antibody against the causative bacterium in body fluids such as a sera and the like. The former method is called an antigen test and the latter is called an antibody test. Both of them are clinically important. As for
Chlamydia pneumoniae
, there is known an antibody test which is carried out by a method in which an antibody is detected by using an elementary body of
Chlamydia pneumoniae.
However, this method has the disadvantage that the elementary body of
Chlamydia pneumoniae
reacts not only with an antibody against
Chlamydia pneumoniae
but also with antibodies against other species of Chlamydia, thus being fairly unspecific. This is because the elementary body of
Chlamydia pneumoniae
contains an antigen which is also present in other species of geneus Chlamydia than
Chlamydia pneumoniae
, that is,
Chlamydia trachomatis
and
Chlamydia psittaci.
As a plasmid which can be used for the expression of a large amount of a protein in
E. coli
, pBBK10MM is known (Japanese Unexamined Patent Publication No. Hei 4-117284). This plasmid can be used for the expression of a fused protein of an anti-allergic peptide with DHFR. The expressed fused protein also maintains the enzymatic activity of DHFR and can therefore be purified easily by utilizing the characteristic properties and activities of DHFR.
Genetic screening has been carried out to diagnose infections. In this screening, the presence of the gene of a microorganism to be detected in a sample is examined using nucleic acid probes and the like.
As for
Chlamydia pneumoniae
, there is known a genetic screening method which is carried out as disclosed in Japanese Unexamined Patent Publication No. Sho 64-500083, U.S. Pat. No. 5,281,518 and WO94/04549.
However, Japanese Unexamined Patent Publication No. Sho 64-500083 and U.S. Pat. No. 5,281,518 only disclose that a chromosomal DNA of
Chlamydia pneumoniae
or a DNA fragment which is obtained by cleaving the chromosomal DNA with a restriction enzyme or the like is used as a probe. The base sequences of these DNA molecules are not determined and the specificity of these probes are therefore unclear. In addition, it is difficult to determine the reaction conditions.
Although WO94/04549 discloses a method using a probe which is hybridized to ribosome RNA or DNA corresponding thereto, the specificity of these probes is not reliable because the homology of ribosomal RNA is relatively high in all organisms.
DISCLOSURE OF THE INVENTION
It is an object of the invention to provide antigenic polypeptides that do not react with antibodies against species of geneus Chlamydia other than
Chlamydia pneumoniae
, such as
Chlamydia trachomatis, Chlamydia psittaci
and the like and which react only with a
Chlamydia pneumoniae
-specific antibody and can thereby detect the
Chlamydia pneumoniae
-specific antibody.
Another object of the invention is to provide a method for synthesizing large amounts of the antigenic polypeptides by using gene recombination techniques.
A further object of the invention is to provide a method for production of an anti-
Chlamydia pneumoniae
-specific antibody, a method and reagents for detection and/or measurement of the anti-
Chlamydia pneumoniae
-specific antibody, and agents for diagnosis of
Chlamydia pneumoniae
infections, all by using said antigenic polypeptides.
A still further object of the invention is to provide probes and primers for detecting and/or measuring specifically
Chlamydia pneumoniae
gene, a method and reagents for detection and/or measurement of
Chlamydia pneumoniae
gene and agents for diagnosis of
Chlamydia pneumoniae
infections, all by using the probes or primers.
An even further object of the invention is to provide antigenic polypeptides for detection of an antibody which reacts with geneus Chlamydia including
Chlamydia pneumoniae, Chlamydia trachomatis, Chlamydia psittaci
and the like.
SUMMARY OF THE INVENTION
The subject matters of the invention are as follows:
(1) A
Chlamydia pneumoniae
antigenic polypeptide, which comprises polypeptide containing a sequence of at least 5 consecutive amino acids in the polypeptide of SEQ ID NO: 1 (hereinafter referred to as “polypeptide A”).
(2) The antigenic polypeptide of (1), wherein said polypeptide A is a polypeptide in which at least one amino acid is deleted from the polypeptide of SEQ ID NO: 1.
(3) The antigenic polypeptide of (1), wherein said polypeptide A is a polypeptide in which at least one amino acid in the polypeptide of SEQ ID NO: 1 is replaced with other amino acid or a polypeptide in which at least one amino acid is added in the polypeptide of SEQ ID NO: 1.
(4) The antigenic polypeptide of (1), wherein said polypeptide A is a polypeptide in which an amino acid or a peptide sequence is bound to a sequence of at least 5 consecutive amino acids in the polypeptide of SEQ ID NO: 1.
(5) The antigenic polypeptide of (1), wherein said polypeptide A is a polypeptide containing the amino acid sequence of SEQ ID NO: 1.
(6) The antigenic polypeptide of (1), wherein said polypeptide A is a polypeptide containing the amino acid sequence of SEQ ID NO: 2.
(7) The antigenic polypeptide of (1), wherein said polypeptide A is a polypeptide containing the amino acid sequence of SEQ ID NO: 5.
(8) A DNA encoding the antigenic polypeptide of any one of (1)-(7), or a DNA complementary thereto.
(9) The DNA of (8), which contains the base sequence of SEQ ID NO: 3.
(10) The DNA of (8), which contains the base sequence of SEQ ID NO: 4.
(11) The DNA of (8), which contains the base sequence of SEQ ID NO: 7.
(12) A recombinant vector carrying the DNA of any one of (8)-(11).
(13) The recombinant vector of (12), which is plasmid pCPN533 &agr; containing the base sequence of SEQ ID NO: 10.
(14) A transformant containing the recombinant vector of (12) or (13).
(15) A method for production of an anti-
Chlamydia pneumoniae
antibody,
wherein the antigenic polypeptide of any one of (1)-(7) is used as an antigen.
(16) A method for detection and/or measurement of an anti-
Chlamydia pneumoniae
antibody, wherein the antigenic polypeptide of any one of (1)-(7) is used as an antigen.
(17) A reagent for detection and/or measurement of an anti-
Chlamydia pneumoniae
antibody, which comprises the antigenic polypeptide of any one of (1)-(7) as an antigen.
(18) A reagent for diagnosis of a
Chlamydia pneumoniae
infection, which comp
Izutsu Hiroshi
Matsumoto Akira
Obara Kazuhiko
Hitachi Chemical Company Ltd.
Stucker Jeffrey
Winkler Ulrike
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