Sputum smear used for acid-fast bacillus microscope examination in proficiency testing activity and preparation method and application thereof
A technology for acid-fast bacilli and proficiency testing, which is applied in the fields of public health, health supervision, health quarantine, and medical technology, and can solve problems such as sample preparation and result judgment difficulties, achieving highly practical results
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[0072] 2.2 Sample preparation
[0073] 2.2.1. Preparation of glass slides
[0074] Soak the new slides in 95% ethanol overnight, take them out and dry them naturally for later use.
[0075] 2.2.2. Preparation method of negative sputum smear
[0076] Negative sample preparation method: take morning sputum from different non-tuberculosis patients (provided by Shanghai International Travel Health Care Center), place it in a sterile sputum specimen collection container, and inactivate bacteria (80°C water bath, 30 minutes to 120 minutes); After natural cooling, put it in a sterile sputum treatment tube, add NaOH-NALC (sodium hydroxide-N acetylcysteine) 2 times the volume of sputum and mix for 20 minutes, then wash with pH6.8 PBS (phosphate buffer solution) ) to 50ml, centrifuge at 3000g (centrifugal force) for 30 minutes, so that the solids in the sputum are concentrated at the bottom of the test tube, discard the supernatant, take the sediment, add PBS equal to the volume of th...
Embodiment 1
[0078] Embodiment 1 Acid-fast bacillus is derived from the preparation method of sputum
[0079] a) Preparation method of 1+ sputum smear
[0080] Take morning sputum from different tuberculosis patients (provided by Shanghai Center for Disease Control and Prevention), put it in a sterile sputum sample collection container, and inactivate bacteria (80°C water bath, 30 minutes); after natural cooling, put it in sterile sputum In the treatment tube, add 2 times the volume of sputum NaOH-NALC and mix for 20 minutes, then dilute to 50ml with PH6.8PBS, centrifuge at 3000g for 30 minutes, so that the tubercle bacteria are concentrated at the bottom of the test tube, discard the supernatant, and add to the sediment Mix 1ml of PBS as a sputum suspension, detect the number of bacteria in it with an acid-fast staining microscope, and dilute with PBS to a final concentration of 2.2×10 4 Strips / ml of sputum dilution, take 30 μl of the above-mentioned sputum dilution and smear it on a gla...
Embodiment 2
[0087] Embodiment 2 acid-fast bacillus is derived from the preparation method of cultivating bacterial colony
[0088] 1. Preparation of BCG suspension
[0089] Get the bacterial colonies of the first two weeks of growth of BCG provided by Shanghai Institute of Biological Products and put them into the bottom of a glass sterilizer, add 100 μl of PBS containing 0.5% Tween-80, and inactivate the bacteria (80°C water bath, 30 minutes ); after natural cooling, insert the grinding rod and twist it to make it cheese-like, dilute the grinding bacteria with normal saline containing 0.5% Tween-80, compare the turbidity with the standard McFarland turbidimetric tube (MacFarland No.1), and prepare 1mg / ml bacterial suspension.
[0090] 2. Preparation of positive sputum smear
[0091] a) Preparation method of 1+ sputum smear
[0092] Take morning sputum from different non-tuberculosis patients (provided by Shanghai International Travel Health Care Center), put it in a sterile sputum sam...
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