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Merck

A novel bedside diagnostic test for methanol poisoning using dry chemistry for formate.

Scandinavian journal of clinical and laboratory investigation (2015-07-24)
Knut Erik Hovda, Gaut Gadeholt, Vladimir Evtodienko, Dag Jacobsen
RESUMEN

The standard diagnostic approach to methanol poisoning is chromatographic measurement of methanol on centrally placed stationary equipment. Methanol poisoning in places where such equipment is unavailable is thus often not diagnosed. Methanol is metabolized to a toxic metabolite, formate; the presence of this compound indicates methanol poisoning. We have developed an enzymatic test for formate and modified it into a portable dry chemistry system that could be used anywhere. The method consists of two enzymatic steps: Formation of NADH from NAD by formate dehydrogenase, and subsequent use of NADH as a reductant of a tetrazolium into a formazan dye that can be quantified photometrically or visually. The photometer gave a good correlation of R(2) = 0.9893 in serum and R(2) = 0.9949 in whole blood, showing an instrumental detection limit of less than 1 mM (4.5 mg/dL). The visual readings showed a correlation of R(2) = 0.8966. Users experienced some difficulty in separating the negative control from the low positives. We have documented the feasibility of an affordable formate strip test for bedside diagnosis of methanol poisoning and for screening of metabolic acidosis of unknown origin. Visual reading is possible, but a reader will improve reliability at lower levels of formate. Future studies are necessary to study the sensitivity and specificity towards other causes of metabolic acidosis and other acids present in human blood.

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Roche
Formate Dehydrogenase, from Candida boidinii
Sigma-Aldrich
Diaphorase from Clostridium kluyveri, recombinant, expressed in E. coli, Animal-component free