Reliable and Rapid Identification of Listeria monocytogenes and Listeria Species by Artificial Neural Network-Based Fourier Transform Infrared Spectroscopy

ABSTRACT Differentiation of the species within the genus Listeria is important for the food industry but only a few reliable methods are available so far. While a number of studies have used Fourier transform infrared (FTIR) spectroscopy to identify bacteria, the extraction of complex pattern information from the infrared spectra remains difficult. Here, we apply artificial neural network technology (ANN), which is an advanced multivariate data-processing method of pattern analysis, to identify Listeria infrared spectra at the species level. A hierarchical classification system based on ANN analysis for Listeria FTIR spectra was created, based on a comprehensive reference spectral database including 243 well-defined reference strains of Listeria monocytogenes, L. innocua, L. ivanovii, L. seeligeri, and L. welshimeri. In parallel, a univariate FTIR identification model was developed. To evaluate the potentials of these models, a set of 277 isolates of diverse geographical origins, but not included in the reference database, were assembled and used as an independent external validation for species discrimination. Univariate FTIR analysis allowed the correct identification of 85.2% of all strains and of 93% of the L. monocytogenes strains. ANN-based analysis enhanced differentiation success to 96% for all Listeria species, including a success rate of 99.2% for correct L. monocytogenes identification. The identity of the 277-strain test set was also determined with the standard phenotypical API Listeria system. This kit was able to identify 88% of the test isolates and 93% of L. monocytogenes strains. These results demonstrate the high reliability and strong potential of ANN-based FTIR spectrum analysis for identification of the five Listeria species under investigation. Starting from a pure culture, this technique allows the cost-efficient and rapid identification of Listeria species within 25 h and is suitable for use in a routine food microbiological laboratory.

[1]  T. Popović,et al.  Probes and polymerase chain reaction for detection of food-borne bacterial pathogens. , 1995, International journal of food microbiology.

[2]  A G Williams,et al.  Effect of sampling procedure and strain variation in Listeria monocytogenes on the discrimination of species in the genus Listeria by Fourier transform infrared spectroscopy and canonical variates analysis. , 1997, FEMS microbiology letters.

[3]  W. Goebel,et al.  Synthetic peptides derived from the Listeria monocytogenes p60 protein as antigens for the generation of polyclonal antibodies specific for secreted cell-free L. monocytogenes p60 proteins , 1994, Applied and environmental microbiology.

[4]  D. Naumann,et al.  Classification and identification of bacteria by Fourier-transform infrared spectroscopy. , 1991, Journal of general microbiology.

[5]  M. Lessing,et al.  Listeria ivanovii infection. , 1994, The Journal of infection.

[6]  W. Goebel,et al.  Detection and Differentiation ofListeria spp. by a Single Reaction Based on Multiplex PCR , 1999, Applied and Environmental Microbiology.

[7]  H. Deneer,et al.  Species-specific detection of Listeria monocytogenes by DNA amplification , 1991, Applied and environmental microbiology.

[8]  F. Allerberger,et al.  Nonhemolytic strains of Listeria monocytogenes detected in milk products using VIDAS immunoassay kit. , 1997, Zentralblatt fur Hygiene und Umweltmedizin = International journal of hygiene and environmental medicine.

[9]  D. A. A. Mossel,et al.  Detection and enumeration of Listeria monocytogenes in foods , 2003 .

[10]  P. Caumette,et al.  Rapid Identification of Listeria Species by Using Restriction Fragment Length Polymorphism of PCR-Amplified 23S rRNA Gene Fragments , 2003, Applied and Environmental Microbiology.

[11]  J. Rocourt,et al.  API Listeria, a new and promising one-day system to identify Listeria isolates , 1992, Applied and environmental microbiology.

[12]  D. Norton Polymerase chain reaction-based methods for detection of Listeria monocytogenes: toward real-time screening for food and environmental samples. , 2002, Journal of AOAC International.

[13]  H. Gremlich,et al.  Infrared and Raman Spectroscopy of Biological Materials , 2000 .

[14]  C. Holt,et al.  Discrimination of species in the genus Listeria by Fourier transform infrared spectroscopy and canonical variate analysis , 1995, Applied and environmental microbiology.

[15]  Siegfried Scherer,et al.  Rapid and Reliable Identification of Food-Borne Yeasts by Fourier-Transform Infrared Spectroscopy , 1998, Applied and Environmental Microbiology.

[16]  W. F. Harrigan Listeria, Listeriosis and Food Safety, 2nd edition , 2001 .

[17]  B. Ninet,et al.  Assessment of the Accuprobe Listeria monocytogenes culture identification reagent kit for rapid colony confirmation and its application in various enrichment broths , 1992, Applied and environmental microbiology.

[18]  Thomas Udelhoven,et al.  Development of a Hierarchical Classification System with Artificial Neural Networks and FT-IR Spectra for the Identification of Bacteria , 2000 .

[19]  D. Volokhov,et al.  Identification of Listeria Species by Microarray-Based Assay , 2002, Journal of Clinical Microbiology.

[20]  D. Naumann,et al.  Prospective Study of the Performance of Vibrational Spectroscopies for Rapid Identification of Bacterial and Fungal Pathogens Recovered from Blood Cultures , 2003, Journal of Clinical Microbiology.

[21]  D B Kell,et al.  Rapid identification of Streptococcus and Enterococcus species using diffuse reflectance-absorbance Fourier transform infrared spectroscopy and artificial neural networks. , 1996, FEMS microbiology letters.

[22]  J. Rocourt,et al.  [Acute purulent Listeria seelingeri meningitis in an immunocompetent adult]. , 1986, Schweizerische medizinische Wochenschrift.

[23]  Clifford Goodman,et al.  Food and Drug Administration Center for Food Safety and Applied Nutrition , 1988 .

[24]  J. McLauchlin,et al.  Listeria ivanovii infection in a patient with AIDS. , 1994, The Journal of infection.

[25]  Harald Labischinski,et al.  Elaboration of a procedure for identification of bacteria using Fourier-Transform IR spectral libraries: a stepwise correlation approach , 1991 .

[26]  L. Cocolin,et al.  Direct Identification in Food Samples of Listeria spp. and Listeria monocytogenes by Molecular Methods , 2002, Applied and Environmental Microbiology.

[27]  C. Lattuada,et al.  Comparison of Nucleic Acid Hybridization Assays and Biochemical Characterization Tests for the Confirmation of Listeria monocytogenes. , 1993, Journal of food protection.

[28]  C. Buchrieser,et al.  The Genus Listeria and Listeria monocytogenes: Phylogenetic Position, Taxonomy, and Identification , 2007 .

[29]  Differentiation of Listeria monocytogenes, Listeria innocua, Listeria ivanovii, and Listeria seeligeri by pulsed-field gel electrophoresis , 1992, Applied and environmental microbiology.

[30]  W. Goebel,et al.  Isolation of catalase-negative Listeria monocytogenes strains from listeriosis patients and their rapid identification by anti-p60 antibodies and/or PCR , 1997, Journal of clinical microbiology.

[31]  S. Scherer,et al.  High incidence of Listeria monocytogenes in European red smear cheese. , 2001, International journal of food microbiology.

[32]  L. Herman,et al.  Repetitive element sequence‐based PCR for species and strain discrimination in the genus Listeria , 1996, Letters in applied microbiology.

[33]  M. Vaneechoutte,et al.  Comparison of PCR-based DNA fingerprinting techniques for the identification of Listeria species and their use for atypical Listeria isolates. , 1998, International journal of systematic bacteriology.

[34]  Siegfried Scherer,et al.  Identification of coryneform bacteria and related taxa by Fourier-transform infrared (FT-IR) spectroscopy. , 2002, International journal of systematic and evolutionary microbiology.

[35]  Siegfried Scherer,et al.  Fourier-Transform Infrared Microspectroscopy, a Novel and Rapid Tool for Identification of Yeasts , 2002, Applied and Environmental Microbiology.

[36]  L. Cocolin,et al.  Temperature gradient gel electrophoresis of the amplified product of a small 16S rRNA gene fragment for the identification of Listeria species isolated from food. , 2000, Journal of food protection.

[37]  C. Addison,et al.  RAPD typing for distinguishing species and strains in the genus Listeria. , 1994, The Journal of applied bacteriology.

[38]  P. Velge,et al.  Hypovirulent Listeria monocytogenes strains are less frequently recovered than virulent strains on PALCAM and Rapid' L. mono media. , 2003, International journal of food microbiology.