STI Panel Real Time PCR Detection (500 test)

STI Panel Real Time PCR Detection (500 test)

Cat. No.: BMG-QP2158-02
Size: 500 tests


Description
 

At present, sexually transmitted diseases (STDs) make up the most common group of infectious diseases that must be reported in the majority of countries. Their incidence is elevated, and more than 20 pathogenic microorganisms are known to be transmitted by sexual contact, with Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, and Mycoplasma (genitalium and hominis) being the most common.

Sexually transmitted diseases (STDs) are a major cause of infertility, long-term disability, ectopic pregnancy, and premature birth.

The product is intended for use by qualified laboratory personnel well trained in nucleic acid amplification techniques and in vitro diagnostic procedures.

Positive results do not rule out co-infection with other pathogens. Negative results do not preclude microbial infection and should not be used as the sole basis for patient management decisions.

Test results must be combined with clinical observations, patient history, and epidemiological information.

Summary and explanation of the test

The STD Panel test is a multiplex PCR test that allows the detection of 8 pathogens associated to sexually transmitted diseases: Mycoplasma hominis, Chlamydia trachomatis, Gardnerella vaginalis, Trichomonas vaginalis, Ureaplasma parvum serovar, Ureaplasma urealyticum, Mycoplasma genitalium, and Neisseria gonorrhoeae


Principle

After DNA isolation, the primers and probes are selected from the regions of targeted conserved regions of the above bacteria. The primer/probe sets are designed for specific detection of above regions are separated into 2 multiplex qPCR setups. An additional primer/probe set to detect human TERT gene (as an internal control) is also included. DNA extracted from sexually transmitted diseases specimens is amplified in a Real-Time PCR Thermocycler (such as Applied Biosystems 7500 Real-Time PCR Instrument).

The fluorescent signal is emitted when the 5’ nuclease active domain of Taq polymerase degrades the probe anneals to target sequence between the forward and the reverse primers.

During each cycle, the fluorescence intensity is increased with the additional reporter dye molecules released from their respective probes. The STD panel Assay also contains a positive control (Plasmid DNA) and a negative control for testing process monitoring.


Reagents and materials provided
 

Item	Component	Quantity		Volume /T
		100 Tests / Box	500 Tests / Box
1	ST PCR Mix I	1.6 mL	5x1.6 mL	15 µL
2	ST PCR Mix II	1.6 mL	5x1.6 mL	15 µL
3	Positive Control	50 μL	250 μL	5 μL
4	Negative Control	50 μL	250 μL	5 μL

Materials may required but not provided

1. Real-Time PCR Thermocycler compatible with JOE (VIC or HEX is equivalent to JOE), FAM and Cy5 channels.
2. Specimen Collection Containers
3. DNA extraction reagent
4. PCR tubes compatible with Thermocycler.
5. 1.5 mL centrifuge tubes (RNase-Free)
6. Vortex
7. High-speed centrifuge
8. Micropipette (0.5-2 µL, 1-10 µL)
9. Pipette tips with filters
10. Personal protective material

Storage instructions

Store the kit frozen at -20±5°C and avoid repeated freeze-thaw cycles (less than 5 frozen-thaw times allowed). The product is valid for 12 months, and the expiry date of the product is shown on the label.

Specimen type and preservation

Specimen type: Dry Swab, serum, whole blood, biopsies, bronchoalveolar lavages (BAL), cerebrospinal fluid, and vitreous humor.
Specimen preservation: process specimens for fungus DNA extraction and nucleic acid detection as soon as possible. Specimens can be stored at 4°C for 24 hours, or frozen at -20°C for longer storage, if not processed immediately. Avoid repeated freeze-thaw cycles during transport and storage of the specimens.

Assay PCR reaction setup (at reagent preparation area)

A. Reagent Preparation
Add 15 μL of ST PCR Mix to each tube or well.

B. Addition of Bacterial DNA
Add 5 μL of bacteria DNA extracts of the specimen or positive/negative control to each PCR tube according to the reaction preparation table below for testing. Then immediately cap the tube tightly, briefly spin the mixture and move it to the amplification area for PCR detection.

• PCR Reaction Preparation

C. PCR amplification (at Amplification Area, use Applied Biosystems 7500 Real-Time PCR System as an example)

1. 1. Place the complete PCR reaction tubes into the fluorescent quantitative PCR analyzer and label positive control, negative control and testing specimen on the Thermocycler software.
2. 2. Enter the PCR reaction volume: 20 μL.
3. 3. Select fluorescence detection channels: JOE, FAM and Cy5 channels. Cy5 is the E gene indicator channel, FAM is the ORF1ab indicator channel, and JOE is the RNase P gene indicator channel (internal control).
4. 4. Select PCR cycle parameter setting following the instructions below, save the file and start the PCR run.
5. 5. Analyze the results according to your thermocycler manufacturer’s instructions. If the positive and negative control samples do not meet their respective requirements, the PCR run must be repeated.

Determination of results

1. The results can only be determined when the results of both controls meet their requirements.
2. Criterion for a positive specimen: a specimen can be reported as positive when FAM, Cy5 and JOE channels are all positive.
3. Criterion for a negative specimen: a specimen can be reported as negative when the JOE channel is positive and the FAM and Cy5 channels are negative

Compatibility of real-time PCR thermocyclers
Product performance

Product analysis performance evaluation results:

1. 1. The analytical sensitivity of this kit is 200 copies/mL.
2. 2. Cross-reaction: no cross-reaction with other pathogens such as .