BD ProbeTec Manual Del Usario página 3

two separate disposable microwells: the Priming Microwell contains the amplification primers, fluorescently-labeled
detector probe, nucleotides and other reagents necessary for amplification, while the Amplification Microwell
contains the two enzymes (a DNA polymerase and a restriction endonuclease) that are required for SDA. The BD Viper
System pipettes a portion of the purified DNA solution from each Extraction Tube into a Priming Microwell to
rehydrate the contents. After a brief incubation, the reaction mixture is transferred to a corresponding, pre-warmed
Amplification Microwell which is sealed to prevent contamination and then incubated in one of the two thermally-
controlled fluorescent readers. The presence or absence of C. trachomatis DNA is determined by calculating the peak
fluorescence (Maximum Relative Fluorescent Units [MaxRFU]) over the course of the amplification process and by
comparing this measurement to a predetermined threshold value.
In addition to the fluorescent probe used to detect amplified C. trachomatis target DNA, a second fluorescently-
labeled oligonucleotide is incorporated in each reaction. The Extraction Control (EC) oligonucleotide is labeled with a
different dye than that used for detection of the C. trachomatis-specific target and is used to confirm the validity of
the extraction process. The EC is dried in the Extraction Tubes and is re-hydrated upon addition of the specimen and
extraction reagents. At the end of the extraction process, the EC fluorescence is monitored by the BD Viper instrument
and an automated algorithm is applied to both the EC and C. trachomatis-specific signals to report specimen results as
positive, negative, or EC failure.
REAGENTS
Each BD ProbeTec CT Q
x
CT Q Amplified DNA Assay Priming Microwells, 12 x 96: each Priming Microwell contains approximately 110 pmol
oligonucleotides, 45 pmol fluorescently-labeled detector probe, 80 nmol dNTPs, with stabilizers and buffer
components.
x
CT Q Amplified DNA Assay Amplification Microwells, 12 x 96: each Amplification Microwell contains approximately
100 units DNA polymerase and 620 units restriction enzyme, with stabilizers and buffer components.
NOTE: Each microwell pouch contains one desiccant bag.
Control Set for the BD ProbeTec CT/GC Q
approximately 2400 copies each of pCTB4 and pGCint3 linearized plasmids in carrier nucleic acid, and 24 CT/GC Q
Negative Controls Tubes containing carrier nucleic acid alone. The concentrations of the pCTB4 and pGCint3 plasmids
are determined by UV spectrophotometry.
CT/GC Q x Swab Diluent for the BD ProbeTec CT/GC Q
2 mL of potassium phosphate/potassium hydroxide buffer with DMSO and preservative.
BD FOX Extraction Tubes for the BD ProbeTec CT/GC Q
approximately 10 mg of iron oxide in a dissolvable film and approximately 240 pmol fluorescently-labeled Extraction
Control oligonucleotide.
Extraction Reagent and Lysis Trough for the BD ProbeTec CT/GC Q
Reagent trough contains approximately 16.5 mL Binding Acid, 117 mL Wash Buffer, 35 mL Elution Buffer, and 29 mL
Neutralization Buffer with preservative; each Lysis Trough contains approximately 11.5 mL Lysis Reagent.
INSTRUMENT, EQUIPMENT AND SUPPLIES
Materials Provided: BD Viper Instrument, BD Viper Instrument Plates, BD Viper Pipette Tips, BD Viper Tip Waste Boxes,
BD Viper Amplification Plate Sealers (Black), BD Viper Lysing Heater, BD Viper Lysing Rack, BD Viper Neutralization
Pouches, Specimen Tubes and Caps for use on the BD Viper (Extracted Mode), Urine Preservative Transport for the
BD ProbeTec Q x Amplified DNA Assays (Q
x
CT/GC Q Amplified DNA Assays, Male Urethral Specimen Collection Kit for the BD ProbeTec CT/GC Q
Assays, Vaginal Specimen Transport for the BD ProbeTec CT/GC Q
Materials Required But Not Provided: Nitrile gloves, 1% (v/v) sodium hypochlorite*, DNA AWAY™, 3% (w/v) hydrogen
peroxide, Chlamydia trachomatis ATCC™ VR-902B (diluted in phosphate buffered saline) or Blackhawk AmpliTrol™
CT/GC.
*Mix 200 mL of bleach with 800 mL of water. Prepare fresh daily.
Storage and Handling Requirements: Reagents may be stored at 2 – 33ºC. Unopened Reagent Packs are stable until
the expiration date. Once a pouch is opened, the microwells are stable for 6 weeks if properly sealed or until the
expiration date, whichever comes first. Do not freeze.
Warnings and Precautions
General:
1. For in vitro Diagnostic Use
2. Pathogenic microorganisms, including hepatitis viruses and Human Immunodeficiency Virus, may be present in
clinical specimens. "Standard Precautions"
items contaminated with blood and other body fluids.
3. For additional specific warnings, cautions and notes specific to the BD Viper, consult the BD Viper System User's
Manual.
Specimen:
4. For collection of endocervical swab specimens, use only the Female Endocervical Specimen Collection Kit for the
BD ProbeTec CT/GC Q
5. For patient-collection and transport of vaginal swabs, use only the Vaginal Specimen Transport for the
BD ProbeTec CT/GC Q
6. For collection of male urethral swab specimens, use only the Male Urethral Specimen Collection Kit for the
BD ProbeTec CT/GC Q
7. For urine specimens, use only the Q
8. Under or over dispensing of urine into Specimen Tubes or the Q
the tubes may also result in liquid overflow on the BD Viper deck, and could cause contamination.
x Reagent Pack contains:
x
Amplified DNA Assays: 24 CT/GC Q
x UPT), Female Endocervical Specimen Collection Kit for the BD ProbeTec
x
Amplified DNA Assays.
x Amplified DNA Assays.
x Amplified DNA Assays.
x UPT or unpreserved (neat) urine.
x Amplified DNA Assays: 48 tubes each containing approximately
x Amplified DNA Assays: 48 strips of 8 tubes, each containing
x Amplified DNA Assays: each 4-cavity Extraction
x Amplified DNA Assay, BD ProbeTec Accessories.
10-13
and institutional guidelines should be followed in handling all
x
2
x
Positive Control Tubes containing
UPT may affect assay performance. Over filling
x
x
Amplified DNA