Rapid Methods for Microbes Identification

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Presentation transcript:

Rapid Methods for Microbes Identification By : Lucia Dhiantika Witasari

RAPID METHODS AND AUTOMATION

A. Immunoassays for Rapid Detection of Pathogens 1. Immunofluorescence Method Commercially available specific fluorescence-conjugated antibodies (against somatic or flagellar antigens of a pathogen) are mixed with an enriched medium suspected to contain the specific pathogen, such as Salmonella (antigen), on a glass slide. 2. RPLA (reversed passive latex agglutination) Method The antibody of a specific toxin is immobilized on latex particles that are then mixed with a sample preparation suspected to contain toxin (antigen) in wells of microtitration plates. 3. Immunoimmobilization Method For motile pathogens such as Salmonella and Esc. coli O157:H7. 4. Enzyme Immunoassay (EIA) or ELISA Methods Salmonella, Lis. monocytogenes, and Cam. jejuni. 5. Magnetic Immunobeads Method to immunocapture the cells (antigen) of a specific pathogen present in a food suspension with the help of magnetic immunobeads. The immunobeads are plastic-covered ferrous beads coated with the specific antibody. Lis. monocytogenes

ELISA

B. Nucleic Acid Probe for Detection of Pathogens 1. Hybridization Method by hybridizing the target nucleic acid sequence to a synthetic oligonucleotide (probes or primers) which is complementary to the target sequence. 2. Polymerase Chain Reaction (PCR) 3. MULTIPLEX PCR (mPCR) 4. REAL-TIME OR QUANTITATIVE PCR (qPCR) 5. NUCLEIC ACID SEQUENCE-BASED AMPLIFICATION (NASBA) By Compton (1991) 6. LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) By Notomi et al. (2000) 7. Oligonucleotide DNA microarray