5'-Atp → Adenosine Monophosphate

HRID 1906361Notes

Reaction equation

HRID 1906361 reaction equation
[P:1]([O:13][CH2:14][C@H:15]1[O:19][C@@H:18]([N:20]2[C:29]3[N:28]=[CH:27][N:26]=[C:24]([NH2:25])[C:23]=3[N:22]=[CH:21]2)[C@H:17]([OH:30])[C@@H:16]1[OH:31])([O:4]P(OP(O)(O)=O)(O)=O)(=[O:3])[OH:2].C1N=C(N)C2N=CN([C@@H]3O[C@@H]4COP(O)(O[C@H]4[C@H]3O)=O)C=2N=1>>[CH:27]1[N:26]=[C:24]([NH2:25])[C:23]2[N:22]=[CH:21][N:20]([C@@H:18]3[O:19][C@H:15]([CH2:14][O:13][P:1]([OH:4])([OH:3])=[O:2])[C@@H:16]([OH:31])[C@H:17]3[OH:30])[C:29]=2[N:28]=1

Participants

3

Reactant 2

HCID 59575'-AtpHCID 6076Camp

Product 1

HCID 6083Adenosine Monophosphate

Conditions

Temperature
-20 °C

Procedure

This step involves incubation of the sample with ATP and calmodulin to generate cAMP. The assay may be initiated at any time within 48 hours of specimen collection, usually at a central laboratory. The assay is carried out in a sterile solution which encourages bacterial viability (e.g. Stainer-Scholte medium, Stainer, D. W. and Scholte, M. J., A simple chemically defined medium for the production of phase I Bordetella pertussis. J. Gen. Microbiol. 63:211-220, 1971) supplemented with 200 nM to 5 mM (e.g. about 1 mM) adenosine triphosphate, 0,05 μM to 5 μM (e.g. about 0.1 μM) calmodulin, and radiolabeled cAMP in 1 nM to 200 nM concentration as appropriate for optimal sensitivity. Alternatively, any medium which permits the preservation and short term survival of Bordetella may be substituted. The assay solution is maintained frozen at about -20° C. until the day of use. The assay solution is thawed. The cap is removed from the transport tube and a measured amount of 0.5 to 2 ml (e.g. about 1 ml) of assay medium is added on top of the transport medium. The swab is not removed. The tube is capped and incubated at 35°-37° C. upright with rotation for 24 to 48 hours.

Workup

  1. customThe assay may be initiated at any time within 48 hours of specimen collection, usually at a central laboratory
  2. concentration63:211-220, 1971) supplemented with 200 nM to 5 mM (e.g. about 1 mM) adenosine triphosphate, 0,05 μM to 5 μM (e.g. about 0.1 μM) calmodulin, and radiolabeled cAMP in 1 nM to 200 nM concentration as appropriate for optimal sensitivity
  3. temperatureThe assay solution is maintained
  4. customThe cap is removed from the transport tube
  5. additiona measured amount of 0.5 to 2 ml (e.g. about 1 ml) of assay medium is added on top of the transport medium
  6. customThe swab is not removed
  7. customThe tube is capped

Sources & evidence

ORD record
ord-87ef315811b2481aaaef9ea12e2296f3
Source dataset
uspto-grants-1987_03
Patent number
US04652521

Reaction notes

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