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Samples were harvested after 7- and 14-days incubation and extracted in lysis buffer (50 mM Tris pH7.4, 150 mM NaCl 2 and 0.01% Tween80) by bead-beating with 0.1 mm zirconia-silica beads (Sigma) in the FastPrep at 6.5 for 3 45 s with careful cooling between bursts

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For vitamin B12 deficiency , treatment varies according to cause and severity: Dietary deficiency : Oral cyanocobalamin supplementation (50150 micrograms daily between meals) is usually effective Pernicious anaemia or malabsorption : Intramuscular hydroxocobalamin injections are required, typically 1 mg three times weekly for two weeks, then maintenance injections every two to three months lifelong Neurological involvement : More intensive initial treatment with 1 mg hydroxocobalamin on alternate days until no further improvement [2] [18] Follow-up should include symptom review and full blood count monitoring after 1-2 months of treatment

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The tool followed the decision tree in Supplementary Fig