the MiL, Inc.

MICROBE INOTECH LABORATORIES, INC.

Microbial Identification Methods

Microbial Identifications

The MiL, Inc. routinely performs rapid microbiology identifications, confirmations, and tracks contaminant sources of all pathogenic, indicator, spoiler organisms, and other

environmental isolates using the latest technologies.

MicroSeq® 16S & 28S rRNA Gene SequencingGenomic DNA is extracted directly from dead or alive bacterial or fungal colonies grown under any conditions. The 16S rRNA gene(for bacterial identifications) or the 28S rRNA gene (for fungal identifications) is amplified using universal primers and termalcyclers. The amplified 16S or 28S rRNA gene is sequenced using dye terminator cycle sequencing chemistry. The sequence reactions are analyzed using automated DNA sequencers and software. Unknown bacterial and fungal samples are identified using the MicroSeq™ microbial identification software and the 16S or 28S rRNA gene sequence database as well as other public databases. Dependant upon the level of confidence, speed, and cost desired, either the entire 1540 base pair 16S or 28S rRNA gene, or a smaller portion of the gene (500 bp) can be sequenced. A printout of the actual sequence data and the identification is included in each report.

qPCRQuantitative real-time polymerase chain reaction (qPCR) is a laboratory technique use to amplify and, at the same time, quantify a targeted DNA molecule and is made possible though the combined use of specialized PCR reagents (e.g., TaqMan) and refined instrumentation. PCR has the ability to amplify specific DNA sequences in an exponential fashion by in vitro DNA synthesis. The PCR technique can be used to detect, identify, and differentiate agents present in either clinical or environmental samples. This technique is particularly useful in environmental microbiology because the population

size (the amount of a particular organism) can be determined and thus population changes can be tracked over time or in response to a change in the environment. The undisputed success of detection assays based on PCR are largely due to its speed in comparison with many conventional diagnostic methods, such as cultures.In addition to the identification methods mentioned above, you may also be interested in these following tests:

DiversiLab™ & “Bacterial Barcodes” – Strain Sub-TypingBased on repetitive sequence-based PCR (rep-PCR), the DiversiLab™ System allows for the amplification of many different sized DNA fragments. These fragments, when separated, then generate “barcodes” that represents the DNA within the non-coding repetitive sequences of the genome. These fingerprints provide strain level discrimination for both bacteria and fungi.

VIDAS® - Automated ELISA Pathogen DetectionVIDAS is a rapid and reliable automated immunoassay system capable of running multpile tests at the same time and is used for the detection of various bacterial and viral pathogens and toxins, such as Salmonella, Campylobacter, Listeria, and many others (additional tests available upon request). VIDAS has been approved for food, pharmaceutical, medical testing.

Biolog®This Automated metabolic characterization system uses a 96-well microtiter plate arrayed with different carbon sources and the microorganism of interest. Each microorganism has unique capacities to oxidize the various carbon sources. Tetrazolium dye in each of the wells turn darker shades of purple as the carbon sources are oxidized by the microorganism. The test yields a characteristic pattern of positive (purple) and negative wells which provide a metabolic signature with the database of interest to determine an identification. Databases include aerobes, anaerobes, gram positive & negative bacteria, yeast, actinomycetes, and lactic acid bacteria.

RiboPrinter®The RiboPrinter® system characterizes bacteria by creating a fingerprint pattern of an isolate’s genetic material. Pure colonies are isolated from culture, inactivated by heat, and loaded into the sample carrier. DNA is extracted from the cell lysate and is digested to completion using an EcoR1 restriction enzyme. The DNA fragments are separated according to molecular size by gel electrophoresis and are transferred to moving Nylon membranes. After denaturation, each membrane is hybridized with chemically labeled rRNA operon probe from E. coli. The membrane is washed and treated with a blocking buffer and an antisulfonated, DNA antibody/alkaline phosphate conjugate. Unbound conjugates are removed and a chemiluminescent substrate is applied. This step makes each electrophoresis band containing genetic information from the rRNA genes visible to a custom CCD camera in the RiboPrinter® system. The camera detects the light intensity of the targeted bands and converts the patterns from the image data and is compared to other known patterns stored in the database to characterize and identify each sample.

Biolog® plate showing positive results (purple).

VITEK® 2 CompactThe VITEK® 2 Compact is a fully automated bacterial identification and antibiotic susceptibility testing system that uses fluorescent technology to provide broad profiles for the reliable identification of the most relevant organisms. With the growing number of clinically significant pathogens rapidly developing resistance to antimicrobials, fast and accurate identification is a must. Turnaround time for test results have substantially decreased when compared to previous iterations of the VITEK system—the average time to result on gram-positive and gram-negative card identifications is 2-8 hours, while results on gram-positive spore forming bacilli are read at 14 hours, and yeast-like organisms are read at 18 hours.

GC-FAMEGas Chromatograph Fatty Acid Methyl Ester (GC-FAME, also known as Cellular Fatty Acid Analysis) allows for bacterial or yeast identification based on the specific fatty acid composition of the cell wall. Fatty acids are extracted from cultured samples and are separated by gas chromatography. A computer generated, unique profile pattern of the extracted fatty acids (9-20 carbon chains long) is compared through pattern recognition programs to our microbial databases of well over 2800 species and subspecies. Our reports include the fatty acid profiles, a listing of the best database matches, along with an assigned statistical probability value indicating the confidence level of the match.

Our Mission

At Microbe Inotech Laboratories, Inc., it is our mission to provide our clients with excellence in microbiology, service, value, and expertise to help them understand and resolve their biological and biochemical problems. We take great pride in our ability to provide you with the microbial knowledge, techniques, and equipment to satisfy your needs.

Need More Information?

Call us today for information on the services we can provide you and/or your business. If you prefer, you can schedule a one-on-one visit to our facilities to discuss your needs and take a tour of our labs.

Personalized customer service and getting accurate and cost-effective information to our clients in a timely manner is our distinction. Questions about test methods or results are answered by our lab staff—all degreed (Ph.D./M.S./B.S.) microbiologists and biochemists.

11754 Westline Industrial DriveSt. Louis, Missouri 63146-3402

Phone: 314-645-2177Fax: 314-645-2544

E-mail: info@microbeinotech.com

the MiL, Inc.

MICROBE INOTECH LABORATORIES, INC.

All trademarks and registered trademarks are the property of their respective companies.

About Microbe Inotech Laboratories, Inc.

Microbe Inotech has over twentyfive years of experience providing microbiological services to our customers. As your full service microbiological lab, we strive to provide excellence in microbiology and service through our five distinct laboratory sectors: environmental, food safety, industrial, pharmaceutical and governmental.

Merging Innovation & Technology

In an effort to solve problems related to microbial detection and identification, Microbe Inotech was founded in 1991 by Dr. Bruce C. Hemming. Dr. Hemming has more than 40 years of experience with research and development in plant pathology and advanced genetics, 10 of which involved microbiological research with Monsanto. For the past two decades, Microbe Inotech Laboratories, Inc. is proud to have offered many solutions to different issues involving microorganisms.

Detection, Identification & Enumeration

At Microbe Inotech Labs, we work with many top companies performing research and development as well as microbial testing and expert witness testimony. Our expansive databases contain profiles of thousands of species of microbes. Let our professionals complete your microbial diagnostic research and assist in the development of product solutions, reducing risk and saving you both time and resources.

We are registered members and participants in the following organizations:

Microbe Inotech Laboratories, Inc.

11754 Westline Industrial Drive

St. Louis, Missouri 63146-3402Phone: 314-645-2177Fax: 314-645-2544

E-mail: info@microbeinotech.com

Visit us online atwww.microbeinotech.com