Agar®SIP7 indicator paper system
(with diphtheria antitoxin to determine the toxigenicity of diphtheria corynebacteria)
APPOINTMENT
The product for in vitro diagnostics is designed to determine the toxigenic properties of Corynebacterium diphtheriae in the immunoprecipitation reaction.
COMPOSITION
System indicator paper Agar®SIP7 diphtheria antitoxin - 1 vial (20,25,30,40 discs).
Indicator paper systems (SIP) for the identification of microorganisms are disks with a diameter of 6 mm made of chromatographic paper containing certain amounts of reagents.
ANALYZED SAMPLES
The research should be carried out with a pure culture, after determining whether it belongs to the genus Corynebacterium (scattering on lamellar media, microscopy of a smear stained by Leffler).
To work, use cultures grown from 18 to 24 hours at a temperature of 37°C on the surface of korinebakagar or weakly alkaline nutrient agar containing 10% horse serum or bovine serum or 5% hemolyzed blood.
METHOD OF APPLICATION
1. Dry the Petri dish with the medium for determining the toxigenicity of diphtheria microbes at a temperature of 37°C for 15 to 20 minutes, turning the cup and lid upside down.
2. After the specified time, place indicator paper discs with diphtheria antitoxin on the surface of the agap with sterile tweezers. Around each disk with antitoxin, form 5 "plaques" from a daily agar culture with a bacteriological loop with a diameter of 2 mm: alternating 2 "plaques" of the control toxigenic strain and 3 experimental "plaques" (from one analysis with suspicious colonies, form at least 2 "plaques" and 1 plaque" from a mixture of 3 to 6 colonies of the same type; the material from this analysis can occupy the places around another disc with antitoxin). All 5 "plaques" should be placed symmetrically around the disk at a distance of 0.6 or 0.7 cm from the edge. The diameter of each "plaque" is 0.7 cm. Up to 4 discs with antitoxin and, accordingly, up to 20 "plaques" with cultures can be placed on one Petri dish.
3. Place the cups with seeds (upside down) in a thermostat and incubate for 24 to 48 hours at a temperature of 37°C.
4. Toxicity should be assessed after 18-24 hours and finally after 48 hours of incubation at a temperature of 37°C.
5. Test cultures are considered toxigenic if the resulting precipitation lines merge at an angle with the precipitation lines of the control toxigenic strain.
The absence of precipitation lines in the experimental cultures after 48 h in the presence of them in the control toxigenic strain indicates the absence of toxigenicity in the studied cultures.
STORAGE
Store at a temperature of 2-8 °C in a place protected from light after each use. Shelf life 1 year.