Development of a PCR-Restriction Fragment Length Polymorphism Protocol for Rapid Detection and Differentiation of Four Cockroach Vectors (Group I “Dirty 22” Species) Responsible for Food Contamination and Spreading of Foodborne Pathogens: Public Health Importance

Abstract

Assessing the adulteration of food products and the presence of filth and extraneous materials is one of the measures that the U.S. Food and Drug Administration (FDA) utilizes in implementing regulatory actions of public health importance. To date, 22 common pest species (also known as the "Dirty 22" species) have been regarded by this agency as the spreaders of foodborne diseases. We have further categorized the Dirty 22 species into four groups: I has four cockroach species, II has two ant species, III has 12 fly species, and IV has four rodent species. The presence of any Dirty 22 species is also considered an indicator of unsanitary conditions in food processing and storage facilities. In this study, we describe the development of a two-step nested PCR protocol to amplify the small subunit ribosomal gene of group I Dirty 22 species that include four cockroach species: Blattella germanica, Blatta orientalis, Periplaneta americana, and Supella longipalpa, along with the development of a PCR-restriction fragment length polymorphism method for rapid detection and differentiation of these violative species. This method will be utilized when the specimen cannot be identified with conventional microscopic taxonomic methods, especially when only small body parts are separated and recovered from food samples for analysis or when these body parts are in a decomposed state. This new PCR-restriction fragment length polymorphism will provide correct identification of group I Dirty 22 species; this information can then be used in regulation and prevention of foodborne pathogens.

Molecular Identification of Isolated Fungi from Unopened Containers of Greek Yogurt by DNA Sequencing of Internal Transcribed Spacer Region

“…Of the various conserved or regulatory gene used as a genetic marker, the rRNA has been proven to be most successful in developing rapid diagnostic methods for the identification of human-pathogenic microorganisms as well as their vectors [ 14 , 15 , 16 ]. As this locus exist in all organisms often with several copies in tandem repeats evolving at slower evolutionary rate, it is characterized as the prime target to establish phylogenetic relationship [ 17 , 18 ].…”

Genetic Characterization of Fungi Isolated from the Environmental Swabs collected from a Compounding Center Known to Cause Multistate Meningitis Outbreak in United States Using ITS Sequencing

“…The rRNA genes are the most abundant housekeeping genes present in all organisms often with several copies evolving at a slower evolutionary rate [ 11 , 12 ]. Thus, this locus has been extensively used as the genetic marker to understand the phylogenetic relationship, and in developing diagnostic assays for rapid detection and differentiation of pathogenic species and their vectors [ 13 , 14 , 15 , 16 , 17 ].…”

Molecular Surveillance of Cronobacter spp. Isolated from a Wide Variety of Foods from 44 Different Countries by Sequence Typing of 16S rRNA, rpoB and O-Antigen Genes

“…Furthermore, multilocus sequence typing (MLST) has been reported suitable for finding genetic polymorphism in microbes with low natural genetic diversity [33,34,35]. A 7-loci based Cronobacter -specific MLST was developed [36], and the MLST was subsequently used to understand genetic diversity of recovered C. sakazakii isolates from PIF, ingredients of PIF, and their production premises [37,38,39,40,41].…”