Isolation and Identification of Nontuberculous Mycobacteria Associated with Tattoo-related Outbreaks

Abstract

Recently, there have been several tattoo-related outbreaks of nontuberculous mycobacterial skin infections in the United States. In an eort to halt the outbreaks and to prevent similar events from occurring, FDA conducted an investigation to determine the source of the contamination. During the investigation, environmental and water samples were collected from tattoo parlors and manufacturers of tattoo ink. These samples were subjected to isolation of mycobacteria at Wadsworth Center of the New York State Department of Health and at FDA PRLSW followed by species identification of the isolates at PRLSW. In order to conduct the investigational studies in a time-sensitive manner, a two-step screening and classification procedure was devised. In this scheme, suspect mycobacterial colonies were screened using multiplex real- time PCR coupled with melting curve analyses specific for the genus Mycobacterium and for dierentiating the species within the M. chelonae-M. abscessus group. Mycobacterial isolates were subsequently identified via sequencing analysis within the coding regions of both 16s rRNA and RNA polymerase subunit beta. In total, 45 colonies of Mycobacterium were isolated and identified as M. chelonae, M. immunogenum, and M. mucogenicum. The isolates from each set of samples contained the corresponding species of Mycobacterium recovered from outbreak patients. Our results suggested that both unsanitary manufacturing processes during production of tattoo ink and the use of non-sterile water for dilution of tattoo ink were possible causes for outbreaks of skin infection in clients of the aected parlors. In addition, the two-step approach taken for screening and identifying mycobacterial colonies in the current study facilitated rapid investigation of tattoo-related outbreaks of nontuberculous mycobacterial infection, thereby enhancing FDAs ability to better protect public health.