Abstract and subjects
The successful isolation of mycolactone in a laboratory or from a clinical sample relies on proper handling and storage of the toxin. Mycolactone is a light-sensitive and an amphiphilic toxin produced by Mycobacterium ulcerans. The biochemistry of the toxin makes it unstable in aqueous matrices such as blood, which causes it to self-aggregate or present in complex with carrier molecules. This biochemistry also impacts the use of the toxin in vitro, in that it tends to aggregate and stick to substrates in an aqueous environment, which alters its physiological presentation and limits its availability in a sample. Glass materials (i.e., tubes, vials, syringes, plates) should be used when possible to avoid loss of mycolactone sticking to plastic surfaces. Dark containers such as amber vials or aluminum-foil wrapped tubes should be used to avoid photodegradation of the toxin upon exposure to light. Sample storage in organic solvents is ideal for mycolactone stability and recovery; however, this is not always amenable as multiple diagnostic assays might be performed on a single sample (such as PCR or ELISA). In these cases, samples can be stored in an aqueous solution containing a small amount of detergent to enhance recovery of the toxin, and in order to avoid aggregation. Therefore, the downstream manipulations should be carefully considered prior to sample collection and storage. Here we present considerations for the optimal handling and storage of mycolactone in order to obtain quality yield of the toxin for various research and diagnostic applications.