Standort in Deutschland, wo man günstige und qualitativ hochwertige Kamagra Ohne Rezept Lieferung in jedem Teil der Welt zu kaufen.
Wenn das Problem der Verringerung der Potenz berührt mich persönlich war ich schockiert, dass das passiert gerade mit mir kamagra Übrigens jeder leisten und gibt eine sofortige Wirkung ohne Hausarbeiten Anwendungen.
Gesellschaft für klinische
Gesellschaft für klinische
Recombinant enzyme immunoassay for the detection
of specific IgM antibodies to chlamydial LPS in human serum
Chlamydiae are gram-negative bacterial pathogens. They have an obli-
gate intracellular life cycle in mucosal surfaces, endothelial cells,
smooth muscle cells, and according to recent findings in certain tis-
sue structures of the central nervous system. Chlamydiae depend on en-
ergy-rich phosphates of their host cells and are therefore energy
The genus chlamydia comprises four species: C. pneumoniae, C. tracho-
matis, C. psittaci,
and C. pecorum
. C. pneumoniae
and C. trachomatis
are obligate pathogens of humans. C. psittaci
is pathogenic for humans
and a variety of animal species. To date, C. pecorum
has been isolated
from animals only. Chlamydia trachomatis
is one of the most frequent sexually transmitted
pathogens worldwide. It causes infections of the urogenital tract and
the eye. In most cases C. trachomatis
infections are asymptomatic.
This results in a variety of chronic diseases, which are sustained by
the ascended and persisting agents. The clinical pictures include in
women: Endometritis, adnexitis, periappendicitis, perihepatitis, and
reactive arthritis. As a consequence of repeated adnexitis the tubes
occlude which results in sterility.
In men the pathogens may ascend to the epididymis (Æ epididymitis) and
into the prostate gland (Æ prostatitis) after incomplete or unsuccess-
ful treatment of urethritis. A reduction of fertility has been discus-
sed in these cases. Furthermore, post-urethritic reactive arthritis is
known in men.
In newborns C. trachomatis
can be transferred during delivery from the
infected birth channel to the infant and can cause neonatal conjuncti-
vitis and/or pneumonia.
infections occur worldwide. In addition to flu-
like illness, the clinical picture includes sinusitis, pharyngitis,
bronchitis, chronic obstructive pulmonary disease, atypical pneumonia,
and reactive arthritis. A causal involvement in infection-conditioned
asthma, sarcoidosis, atherosclerosis, acute myocardial infarction,
stroke, multiple sclerosis, and late-onset of Alzheimer’s disease has
been discussed. Chlamydia psittaci
infects birds and mammals, which may transmit the
pathogen to humans. The resultant frequently severe pneumonias may
lead to life-threatening conditions if no targetted antibiotic inter-
vention is started rapidly.
The usual laboratory methods for the detection of chlamydia infections
include antigen and antibody determinations. The antigen detection
(IFA, ELISA, PCR) is of value for the diagnosis of peripherally local-
ized infections. In ascended courses of disease the detection of the
pathogen is limited and has to be complemented by serology. Sero-
logical methods include complement fixation test, IFA, MIF, genus- and
species-specific ELISA systems.
Chlamydiae contain as common immunodominant antigen the lipopolysac-
charide (LPS), to which the first immune reaction is directed. The
corresponding antibodies are already detectable within a few days af-
ter infection and, thus, allow early diagnosis. The use of paired sera
enables the discrimination of current infections, reinfections, and
reactivations (defined titer increases) from chronic persistent ones
(constant antibody titers). Because of the limited persistence of the
LPS antibodies after successful eradication of the pathogens, the dia-
gnosis of current infections is obviously not influenced by past in-
The Chlamydien-IgG-, IgA-, and IgM-rELISA medac are based upon a
molecularly defined, genetechnologically produced antigen. It is an
exclusively chlamydia-specific fragment from the LPS which has not
been found in any other bacterial LPS. The comparison of sera from
blood donors with sera from patients with urogenital or respiratory
infections has shown, that antibodies to chlamydial LPS are detected
more frequently in patients than in blood donors.
As the reaction is genus-specific, the test result will not differen-
tiate betweeen the chlamydia species. However, as the clinical picture
already will hint to the suspected chlamydia species and additionally
all the chlamydiae show an equal sensitivity towards specific antibi-
otics, the genus-specific reactivity does not hamper the consequences.
The plate is coated with chlamy-dia-specific recombinant LPS frag-ment.
The chlamydia-specific LPS antibo-dies from the specimen bind to the antigen.
Peroxidase-conjugated anti-human IgM antibodies bind to the IgM an-tibodies (P = peroxidase).
Incubation with TMB-substrate (*).
The reaction is stopped by the ad-
dition of sulfuric acid. The ab-
sorption is read photometrically.
Advantages of the test
Chemically defined, chlamydia-specific antigen.
The breakable microwell strips permit efficient use of the test.
Suitable for automation on open ELISA devices.
Cat. no.: 485-TMB
Microplate: 12 x 8 wells (with frame and desiccant vacuum sealed in aluminium bag), breakable, U-form, coated with chlamydia-spe-cific, recombinant LPS fragment and NBCS, ready to use.
Negative control: 1 vial with 1.5 ml, human serum, ready to use,
stained blue, contains NBCS, phenol, ProClin™ 300 and gentamycin sulfate.
Positive control: 1 vial with 1.5 ml, human serum, ready to use,
stained blue, contains BSA, phenol, ProClin™ 300 and gentamycin sulfate.
Wash buffer: 1 bottle with 100 ml PBS/Tween (10x), pH 7.2-7.4, con-
Sample diluent: 1 bottle with 110 ml PBS/Tween/NBCS, pH 7.0-7.2,
ready to use, stained blue, contains ProClin™ 300.
Conjugate: 4 vials with 4.5 ml each, goat anti-human IgM, HRP-conjugated, ready to use, stained red, contains BSA, phenol, Pro-
TMB-substrate: 1 vial with 10 ml, ready to use.
Stop solution: 2 vials with 11 ml each, 0.5 M sulfuric acid (H2SO4), ready to use.
IgG/Rf-absorbent: 1 vial with 4 ml, goat anti-human IgG antiserum, ready to use, contains < 0.1 % sodium azide.
STORAGE AND STABILITY
Do not use the reagents after the expiry date.
REAGENTS AND MATERIALS REQUIRED BUT NOT PROVIDED
2.1. Water for injection (H2O redist.). Use of deionised water can
2.2. Adjustable micropipettes. 2.3. Clean glass or plastic containers for dilution of wash buffer
2.4. Suitable device for microplate washing (e.g. multistepper or
2.5. Incubator for 37 °C. 2.6. Microplate reader with filters for 450 nm and 620 - 650 nm.
PREPARATION OF THE REAGENTS
Before starting the test procedure all kit components must be equili-
brated to room temperature (RT).
Calculate the number of wells required.
After each removal of wells the aluminium bag has to be tightly
resealed together with the desiccant. Storage and stability of the wells are indicated under point 1.
Mix one volume of wash buffer (10x) with nine volumes of water for injection (e.g. 50 ml wash buffer (10x) with 450 ml water). 10 ml of diluted wash buffer are needed for eight wells.
Crystals in the wash buffer (10x) have to be dissolved by warm-
ing (max. 37 °C) and/or stirring at RT.
Do not mix test specific reagents (microplate, controls, conjugate)
from different kit lots. In contrast to that, sample diluent, wash
buffer, TMB-substrate, IgG/Rf-absorbent and stop solution are gener-
ally exchangeable in all Chlamydia and Mycoplasma-ELISA medac.
Reagents from other manufacturers must not be used in general.
Valid and reproducible results are only obtained if the test proce-
dure is precisely followed.
4.1. The test is suitable for serum samples, but not for plasma.
4.2. Pretreatment of sera, e.g. inactivation, is not necessary. How-
ever, they should neither be contaminated with microorganisms nor contain red blood cells.
4.3. The sera are employed at a final dilution of 1:50 after IgG/Rf
5.A. IgG/RF ABSORPTION
The controls are ready to use (no absorption necessary).
The volumes indicated in the following are for single determi-
5.A.1. Serum: 10 µl serum are diluted with 240 µl sample diluent (di-
5.A.2. Absorption: 30 µl IgG/Rf-absorbent and 30 µl diluted serum are
mixed (dilution 1:50) and incubated for 15 min at RT.
Alternative: The absorption can be performed overnight at 2 - 8 °C.
5.B.1. Cut the aluminium bag above the zip fastener and take out the
required number of microplate wells (see 3.1.).
Microplate wells are ready to use and do not have to be pre-
5.B.2. Pipette 50 µl of sample diluent into the well A1 as blank (see
6.A.), 50 µl of the negative control in duplicate, and each 50 µl of the positive control and the diluted patient samples for
If necessary, the microplate wells can be kept in a humid
chamber up to 30 min at 2 - 8 °C before proceeding.
5.B.3. Incubate the microplate wells for 60 min (± 5 min) at 37 oC
(± 1 °C) in a humid chamber or sealed with incubation cover foil.
5.B.4. After incubation wash the microplate wells three times with
each 200 µl wash buffer per well. Pay attention that all wells are filled. After washing tap microplate wells on filter pa-per.
Do not allow the wells to dry out! Proceed immediately!
5.B.5. Add conjugate (coloured red) to each well.
50 µl of conjugate have to be pipetted into the wells if the
test is done manually.
When working with automated instruments, 60 µl of conjugate
have to be pipetted into each well due to a higher evaporation
in the incubation chambers of the automates.
The principal suitability of the test for automation could be
shown during validation. Nevertheless we recommend to verify
the compatibility with the automate employed.
5.B.6. Incubate the microplate wells again for 60 min (± 5 min) at
37 oC (± 1 °C) in a humid chamber or sealed with incubation cover foil.
5.B.7. After incubation wash microplate wells again (see 5.B.4.).
5.B.8. Add 50 µl of TMB-substrate to each well and incubate the mi-
croplate wells for 30 min (± 2 min) at 37 oC (± 1 °C) in a hu-mid chamber or sealed with incubation cover foil in the dark. Positive samples turn blue.
5.B.9. Stop the reaction by adding 100 µl of stop solution to each
Clean microplate wells from underneath before the photometric reading
and take care that there are no air bubbles in the wells.
The reading should be done within 15 min after adding the stop solu-
TABLE FOR THE IgG/Rf-ABSORPTION
5.D. TABLE FOR THE TEST PROCEDURE
Incubate for 60 min at 37 °C, wash 3 x with 200 µl wash buffer
50/60 µl*) 50/60 µl*) 50/60 µl*) 50/60 µl*)
Incubate for 60 min at 37 °C, wash 3 x with 200 µl wash buffer
Incubate for 30 min at 37 oC in the dark
Photometric reading at 450 nm (ref. 620 - 650 nm)
*) manual/automatic procedure (see 5.B.5.) 6.A. CALCULATION OF RESULTS (VALIDITY)
Read OD values at 450 nm (reference wavelength 620 - 650 nm).
Subtract the OD value of the blank (well A1) from all other OD values.
The OD value of the blank has to be < 0.100.
The mean OD value of the negative control
has to be < 0.200.
The OD value of the positive control
has to be > 0.800.
∗ Cut-off = Mean OD value of the negative control + 0.370
Grey zone = Cut-off ± 15 %
Repeat the run if the results do not meet the specification.
6.B. EVALUATION OF RESULTS
With the Chlamydien-IgM-rELISA medac the sera have to be titrated if
endtiter determination is desired.
6.C. INTERPRETATION OF THE RESULTS
6.C.1.CRITERIA FOR SIGNIFICANT TITER INCREASES
For the diagnosis of current, fresh infections, reinfections, reacti-
vations (significant titer increases) and discrimination from persis-
tent infections (constant antibody titers) we principally recommend
the use of paired sera. They should be obtained 10 - 14 days apart.
The following criteria have been described as a possibility for the
assessment of significant titer increases (Persson et al., Verkooyen
specific IgG or
IgA antibody titers
specific IgG and
IgA antibody titers
SPECIFIC IgM INTERPRETATION
tion. Retest IgM after 10 - 14 days and test for IgA and IgG.
dia IgM antibodies. In cases of jus-tified clinical suspicion test for IgA and IgG.
The IgM results should always be interpreted in connection with IgG and/or IgA, the clinical picture and additional diagnostic parameters.
Samples with OD values within the grey zone should be retested together with a fresh specimen taken 14 days later in order to determine a titer change.
High concentrations of hemoglobin and of bilirubin in serum do not have an influence on the results.
In rare cases, high lipid concentrations in serum may influence the test results.
Cross-reactivities with antibodies to parvovirus B19 cannot be excluded in individual cases.
In cases of fresh acute chlamydial infections the serological anti-
body results may be negative despite clinics and positive antigen de-
tection. If a serological confirmation of a positive antigen result
or if a follow-up is desired we recommend to test after 10 - 14 days
We determined the following performance characteristics during the
diagnostic evaluation. 7.A. PREVALENCE
Sera from various patients cohorts (culture-positive, -negative STD
patients, prostitutes, infertile females, infertile males, patients
with rheumatic diseases, patients with chronic obstructive respira-
tory diseases [COPD]), and controls (pregnant women, 2 blood donor
cohorts) were investigated for the determination of LPS antibody
Culture-positive STD patients
Culture-negative STD patients
Patients with rheumatic diseases
Blood donor group 1
Blood donor group 2
NC = negative control; BC = weak positive control (not included in the kit); PC = positive control
GENERAL HANDLING ADVICES
Do not exchange the vials and their screw caps in order to avoid cross contamination.
The reagents have to be sealed immediately after use in oder to avoid evaporation and microbial contamination.
After use, the reagents have to be stored as indicated to guar-antee the shelf life.
After use, all components of the testkit should be stored in the orginal package, in order to avoid mixing up the reagents of other test systems or lots (see also 3.).
HEALTH AND SAFETY INFORMATION
The local occupational safety and health regulations have to be regarded.
Reagents of human origin have been tested and found to be nega-tive for HBsAg, for antibodies to HIV-1/2 and to HCV. Neverthe-less, it is strongly recommended that these materials as well as those of animal origin (see kit contents), should be handled as potentially infectious and used with all necessary precautions.
Residues of chemicals and preparations are generally considered as hazardous waste. The disposal of this kind of waste is regulated through national and regional laws and regulations. Contact your lo-cal authorities or waste management companies which will give advice on how to dispose hazardous waste.
Date of issue: 01.07.2008
Balin, B.J., Gérard, H.C., Arking, E.J., Appelt, D.M., Branigan,
P.J., Abrams, J.T., Whittum-Hudson, J.A., Hudson, A.P.: Identifica-
tion and localization of Chlamydia pneumoniae
in the Alzheimer’s
brain. Med. Microbiol. Immunol. 187, 23-42 (1998).
Brade, L., Holst, O., Kosma, P., Zhang, Y.X., Paulsen, H., Krausse,
R., Brade, H.: Characterization of murine monoclonal and murine, rab-
bit, and human polyclonal antibodies against chlamydial lipopolysac-
charide. Infect. Immun. 58, 205-213 (1990).
Brade, L., Brunnemann, H., Ernst, M., Fu, Y., Kosma, P., Näher, H.,
Persson, K., Brade, H.: Occurence of antibodies against chlamydial
lipopolysaccharide in human sera as measured by ELISA using an arti-
fical glycoconjugate antigen. FEMS Immunol. Med. Microbiol. 8, 27-42
Diedrichs, H., Schneider, C.A., Scharkus, S., Pfister, H., Erdmann,
E.: Prävalenz von Chlamydien-Antikörpern bei Patienten mit koronarer
Herzerkrankung. Herz/Kreisl. 29, 304-307 (1997).
Elkind, M.S., Lin, I.F., Grayston, J.T., Sacco, R.L.: Chlamydia pneu-
and the risk of first ischemic stroke. The Northern Manhattan
stroke study. Stroke 31, 1521-1525 (2000).
Falck, G., Gnarpe, J., Gnarpe, H.: Persistent Chlamydia pneumoniae
infections in a Swedish family. Scand. J. Infect. Dis. 28, 271-273
Gérard, H.C., Schumacher R.H., El-Gabalawi, H., Goldbach-Mansky, R.,
Hudson, A.P.: Chlamydia pneumoniae
present in the human synovium are
viable and metabolically active. Microbiol. Pathol.29, 17-24 (2000).
Grayston, J.T., Wang, S.P., Kuo, C.C., Campbell L.A.: Current knowl-
edge of Chlamydia pneumoniae
, strain TWAR
, an important cause of
pneumonia and other respiratory diseases. Eur. J. Clin. Microbiol.
Infect. Dis. 8, 191-202 (1989).
Gupta, S. Leatham, E.W., Carrington, D., Mendall, M.A., Kaski, J.C.,
Camm, A.J.: Elevated Chlamydia pneumoniae
events, and azithromycin in male survivors of myocardial infarction.
Circulation 96, 404-407 (1997).
Hahn, D.L., Peeling, R.W., Dillon, E., McDonald, R., Saikku, P.: Se-
rologic markers for C. pneumoniae
in asthma. Ann. Allergy Asthma Im-
munol. 84, 227-233 (2000).
Köhler, M., Jendro, C.: Bedeutung der Persistenz von Chlamydia tra-
im Gelenk für die Pathogenese der Chlamydien-induzierten
Arthritis unter Berücksichtigung der therapeutischen Implikationen.
Akt. Rheumatol. 22, 170-175 (1997).
Land, J.A., Evers, J.L., Goossens, J.V.: How to use Chlamydia anti-
body testing in subfertility patients. Hum. Reprod. 13, 1094-1098
Maass, M., Bartels, C., Engel, P., Mamat, U., Sievers, H.H.: Endovas-
cular presence of viable Chlamydia pneumoniae
is a common phenomenon
in coronary artery disease. JACC 31, 827-32 (1997).
Morré, S.A., De Groot, C.J., Killestein, J., Meijer, C.J., Polman,
C.H., Van der Falk, P., Van den Brule, A.J.: Is Chlamydia pneumoniae
present in the central nervous system of multiple sclerosis patients?
Ann. Neurol. 48, 399 (2000).
Paavonen, J.: Chlamydia trachomatis
: A major cause of mucopurulent
cervicitis and pelvic inflammatory disease in women. In: Sexually
Transmitted Diseases. Advances in Diagnosis and Treatment. Curr.
Probl. Dermatol. Elsner. P., Eichmann, A. (eds.). Basel, Karger, 24,
Patton, D., Askienazy-Elbhar, M., Henry-Suchet, J., Campbell, L.A.,
Cappucio, A., Tannous, W., Wang, S.P., Kuo, C.C.: Detection of Chla-
in fallopian tube tissue in women with post-
infectious tubal infertility. Am. J. Obstet. Gynecol. 171, 95-101
Persson, K., Haidl, S.: Evaluation of a commercial test for antibod-
ies to the chlamydial lipopolysaccharide (Medac) for serodiagnosis of
acute infections by Chlamydia pneumoniae (TWAR)
and Chlamydia psit-
APMIS 108, 131-138 (2000).
Petersen, E.E., Clad, A.: Genitale Chlamydieninfektionen. Deutsches
Ärzteblatt 92, A-277-282 (1995).
Saikku, P.: Diagnosis of acute and chronic Chlamydia pneumoniae
fections. In: Orfila, J. et al. (eds): Chlamydial Infections. Proc.
Eighth Int. Symp. on Human Chlamydial Infect. Societá Editrice Escu-
lapio, Bologna, Italy, p. 163-170 (1994).
Schachter, J.: The intracellular life of Chlamydia. Curr. Top. Micro-
biol. Immun. 138, 109-39 (1988).
Schmitz, F.J., Küppers, B., Reinartz, R., Vossel, R., Schuppe, D.,
Bielfeld, D., Heinz, H.P.: Prevalence of Chlamydia trachomatis
gen and Chlamydia antibodies in prostitutes, infertile female and in-
fertile male patients – Comparison of different methods. In: Stary,
A. (ed.): Proc. Europ. Soc. Chlamydia Res., Societá Editrice
Esculapio, Bologna, Italy, p. 339 (1996).
Sriram, S., Stratton, C.W., Yao S.: Chlamydia pneumoniae
in the central nervous system in multiple sclerosis. Ann. Neurol. 46,
Verkooyen, R.P., van Lent, N.A., Mousavi Joulandan, S.A., Snijder,
R.J., van den Bosch, J.M., van Helden, H.P., Verbrugh, H.A.: Diagno-
sis of Chlamydia pneumoniae
infection in patients with chronic ob-
structive pulmonary disease by micro-immunofluorescence and ELISA. J.
Med. Microbiol. 46, 959-964 (1997).
Verkooyen, R.P., Willemse, D., Hiep-van Casteren S.C.A.M., Mousavi
Joulandan, S.A., Snijder, R.J., van den Bosch, J.M.M., van Helden,
H.P.T., Peeters, M.F., Verbrugh, H.A.: Evaluation of PCR, culture,
and serology for diagnosis of Chlamydia pneumoniae
tions. J. Clin. Microbiol., 36, 2301-2307 (1998).
Ward, M.E.: The immunobiology and immunopathology of chlamydial in-
fections. APMIS 103, 769-796 (1995).
Weström, L.V.: Chlamydia and its effect on reproduction. J. Brit.
Fertil. Soc. 1, 23-30 (1996).
Wollenhaupt, J., Zeidler, H.: Klinik, Diagnostik und Therapie der
Chlamydien-induzierten Arthritis. Akt. Rheumatol. 22, 176-182 (1997).
REPORTABLE IN THE SUPREME COURT OF INDIA CIVIL APPELLATE JURISDICTION CIVIL APPEAL NO. 10209 OF 2011 (Arising out of SLP (C) No.2798 of 2010) J U D G M E N T P. Sathasivam, J. This appeal raises an important question as to the interpretation of Section 21 of the Legal Services Authorities Act, 1987 (in short ‘the Act’). The question posed for consideration is that w
Canadian Federation of Students House of Commons Standing Committee on Finance August, 2001 123456789012345678901234567890121234567890123456789012345678901212345678901234567890123456789012123456789012345678901234567890123456789012345678901234567890121234567890123456789012345678901212345678901234567890123456789012123456789012345678901234567890123456789012345678901234567890121234567890