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Nucleic Acid Detection Kit based on Enzymatic Probe Isothermal Amplification (EPIA) for Candida Albicans

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Category:Gastrointestinal diseases

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Nucleic Acid Detection Kit based on Enzymatic Probe Isothermal Amplification (EPIA) for Candida Albicans

This kit is intended for the in vitro qualitative detection of the nucleic acid of Candida tropicalis in genitourinary tract samples or clinical sputum samples.
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Introduction

Packing specification

50 tests/kit

 

Intended use
This kit is intended for the in vitro qualitative detection of the nucleic acid of Candida tropicalis in genitourinary tract samples or clinical sputum samples.
The test result of this kit is not the only indicator for the diagnosis of Candida albicans infection in patients. It must be combined with the patient’s clinical characteristics, imaging examinations, and other laboratory test data to correctly identify whether the fungus is infection or colonization and to formulate a reasonable treatment protocol to make the anti-fungal treatment be safe and effective.

 

Main components

 

Sample collection
1. Genitourinary tract swab
2. Sputum

 

Sample processing

1. Genitourinary tract swab

1.1 Add 1-3mL of sterile normal saline to the sterile tube containing the swab specimen, and shake and mix thoroughly. Squeeze the cotton swab and discard it.

1.2 Pipette 1mL of liquid to a 1.5mL RNase-free & DNase-free centrifuge tube (if there are too many secretions, take 0.2mL only), centrifuge at 12000rpm for 3min, discard the supernatant and keep the precipitate.

1.3 Add 180μL of lysozyme buffer (dilute the lysozyme to 20mg/mL with lysozyme diluent) to the precipitate, shake and mix, and treat at 37°C for 30min.

1.4 Take a 1.5mL RNase-free & DNase-free centrifuge tube, and add 200μL of positive control and negative control in sequence. Add 3μL of internal reference control to the sample to be tested, the positive control, and the negative control in sequence, and use the recommended nucleic acid extraction reagents for subsequent sample DNA extraction. Please strictly follow the instructions for use for specific steps. Use DNase and RNase-free water for elution. The recommended elution volume is 100μL.

1.5 After heating the extracted sample DNA at 95°C for 3 minutes, immediately place it in ice bath for 2 minutes and then load the sample to the instrument for detection; if the sample is stored below -18°C, before the detection, heat the sample DNA at 95°C for 3 minutes, immediately place it in ice bath for 2 minutes and then load the sample to the instrument for detection. Avoid repeated freezing and thawing.

Note: the sample DNA that has been processed by heating and ice bath should be immediately pipetted and loaded on the instrument for detection; or immediately pipetted and then loaded on the instrument for detection after being placed at 2-8°C for no more than 4h.

 

2 Respiratory sputum

2.1 Add 4 times volume of 4% NaOH to 1-3mL sputum, shake and mix well. Then incubate at 37°C for 30min to liquefy, and centrifuge at 13000rpm for 15min. Discard the supernatant, add 1mL sterile normal saline to the precipitate and shake well. Centrifuge at 13,000 rpm for 10 minutes, discard the supernatant, and repeat again. Add 180μL lysozyme buffer, shake and mix well, and treat at 37°C for over 30 minutes.

2.2 Take a 1.5mL RNase-free & DNase-free centrifuge tube, and add 200μL of positive control and negative control in sequence. Add 3μL of internal reference control to the sample to be tested, the positive control, and the negative control in sequence, and use the recommended nucleic acid extraction reagents for subsequent sample DNA extraction. Please strictly follow the instructions for use for specific steps. Use DNase and RNase-free water for elution. The recommended elution volume is 100μL.

2.3 After heating the extracted sample DNA at 95°C for 3 minutes, immediately place it in ice bath for 2 minutes and then load the sample to the instrument for detection; if the sample is stored below -18°C, before the detection, heat the sample DNA at 95°C for 3 minutes, immediately place it in ice bath for 2 minutes and then load the sample to the instrument for detection. Avoid repeated freezing and thawing.

Note: the sample DNA that has been processed by heating and ice bath should be immediately pipetted and loaded on the instrument for detection; or immediately pipetted and then loaded on the instrument for detection after being placed at 2-8°C for no more than 4h.

 

Applicable instruments and equipment
ABI 7500 fluorescence quantitative PCR instrument, and Hongshi SLAN-96P automatic medical PCR analysis system

Key words:
Nucleic Acid
EPIA
Candida Albicans
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