Hc2 CT-ID
DNA Test
An in vitro nucleic Acid
Hybridization Microplate Assay with signal Amplification using
Microplate chemiluminescene for the Qualitative Detection of
Chlamydia trachomatics (CT) DNA in cervical Specimen’s.
SUMMARY AND EXPLANATION
Chlamydiae are gram negative organisms with a two-phase life cycle
comprising morphologically distinct infections and reproductive
forms. The Chlamydia trachomatis genome is relatively small,
measuring approximately base pairs. Once inside a host cell,
elementary bodies assemble into membrane bound vacuoles to produce
the metabolically active chlamydial reproductive forms. Chlamydiae
have a membrane bound, genus-specific lipopolysaccharide that has
served as a source of antigen for the production of diagnostic
antibodies.
Conventional methods for the direct detection of Chlamydia
trachomatis in clinical specimens include iodine or giemsa
staining of the organism followed by microscopic evaluation or the
more sensitive use of direct fluorescent antibody staining. The
most widely accepted procedure for Chlamydia detection is the
infection of McCoy cells in cell culture. Optimal cell culture has
excellent sensitivity and specificity for the detection of
Chlamydia, but is a complex, expensive and time-consuming
procedure. Nucleic acid tests are also available for the detection
of a variety of Chlamydia targets, including chromosomal DNA, Mrna,
and the cryptic plasmid common to the vast majority of Chlamydia
trachomatis strains.
PRINCIPLE OF THE PROCEDURE
The hc2 CT-ID DNA Test using Hybrid Capture 2 technology is a
nucleic acid hybridization assay with signal amplification that a
specific Chlamydia RNA probe. The resultant RNA:DNA hybrids are
captured onto the surface of a microplate well coated with
antibodies specific for RNA:DNA hybrids. Multiple conjugated
antibodies bind to each captured hybrid resulting in substantial
signal amplification. As the substrate is cleaved by the bound
alkaline phosphatase, light is emitted, which is measured as
relative units. On a luminometer.
The CT probe contains a probe mixture specifically chosen to
eliminate or minimize cross-reactivity with DNA sequences from
human cells, other bacterial species or Chlamydia species other
than Chlamydia trachomatis. High volume sample-throughput testing
with the hc2 CT-ID DNA Test can be performed utilizing a general
use automated pipetting and dilution system referred to as the
rapid capture system. To enable high volume sample-throughput
testing, all the procedural steps of the assay are performed by
the RCS, with the exception of specimen denaturation,
chemiluminescent signal detection, and result reporting.
REAGENTS
AND MATERIALS PROVIDED
There are 96 tests in one hc2 GC-ID
DNA test kit . the number of patient results will vary, depending
on the number of uses per kit.
1 use = 88 patient results
2 use = 80 patient results
3 use = 72 patient results
4 use = 64 patient results
1 * 0.35 ml Indicator Dye
INDIC : Contains 0.05% w/v sodium azide.
1 * 50 ml Denaturation
Reagent REAG DENAT: Dilute sodium hydroxide (NaOH)
solution.
1 * 5 ml Probe Diluent
DIL PROBE: Buffered solution with 0.05% w/v sodium
azide.
1 * 200 µl CT Probe
PROBE CT: CT RNA probe in buffered solution.
1 * 2 ml Negative
Control CONTROL - : Carrier DNA in specimen Transport
Medium (STM) with 0.05% w/v sodium azide.
1 * 1 ml CT Positive
Calibrator (PC) CAL CT + : 1.0 PG/ML cloned CT DNA and
carrier DNA in STM with 0.05% w/v sodium azide.
1 * 1 ml Quality
Control GC (QC CT) QC CT : 5.0 pg.ml cloned CT DNA and
carrier DNA in STM with 0.05% w/v sodium azide.
1 * 1 ml Quality
Control GC (QC GC) QC GC : 5.0pg/ml cloned GC DNA and
carrier DNA in STM with 0.05% w/v sodium azide.
1 each Capture
Microplate PLATE CAPTURE : Coated with anti-RNA:DNA
hybrid antibodies.
1 * 12 ml Detection
Reagent 1 REAG DET 1 : Alkaline phosphatase-conjugated
antibodies to RNA:DNA hybrids in buffered solution with 0.05% w/v
sodium azide.
1 * 12 ml Detection
Reagent 2 REAG DET 2 : CDP-Star with Emerald II.
1 * 100 ml Wash Buffer
Concentrate BUF WASH X 30 : Contains 1.5% w/v sodium
azide.
