Magnetic separation techniques in diagnostic microbiology

The principles of magnetic separation aided by antibodies or other specific binding molecules have been used for isolation of specific viable whole organisms, antigens, or nucleic acids. Whereas growth on selective media may be helpful in isolation of a certain bacterial species, immunomagnetic separation (IMS) technology can isolate strains possessing specific and characteristic surface antigens. Further separation, cultivation, and identification of the isolate can be performed by traditional biochemical, immunologic, or molecular methods. PCR can be used for amplification and identification of genes of diagnostic importance for a target organism. The combination of IMS and PCR reduces the assay time to several hours while increasing both specificity and sensitivity. Use of streptavidin-coated magnetic beads for separation of amplified DNA fragments, containing both biotin and a signal molecule, has allowed for the conversion of the traditional PCR into an easy-to-read microtiter plate format. The bead-bound PCR amplicons can also easily be sequenced in an automated DNA sequencer. The latter technique makes it possible to obtain sequence data of 300 to 600 bases from 20 to 30 strains, starting with clinical samples, within 12 to 24 h. Sequence data can be used for both diagnostic and epidemiologic purposes. IMS has been demonstrated to be a useful method in diagnostic microbiology. Most recent publications describe IMS as a method for enhancing the specificity and sensitivity of other detection systems, such as PCR, and providing considerable savings in time compared with traditional diagnostic systems. The relevance to clinical diagnosis has, however, not yet been fully established for all of these new test principles. In the case of PCR, for example, the presence of specific DNA in a food sample does not demonstrate the presence of a live organism capable of inducing a disease. However, all tests offering increased sensitivity and specificity of detection, combined with reduced time of analysis, have to be seriously evaluated.

[1]  A. Deelder,et al.  Quantitative detection of schistosomal circulating anodic antigen by a magnetic bead antigen capture enzyme-linked immunosorbent assay (MBAC-EIA) before and after mass chemotherapy. , 1992, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[2]  T. Lien,et al.  Immunomagnetically Captured Thermophilic Sulfate-Reducing Bacteria from North Sea Oil Field Waters , 1992, Applied and environmental microbiology.

[3]  E. Beutler,et al.  Interference of heparin with the polymerase chain reaction. , 1990, BioTechniques.

[4]  R. Lanciotti,et al.  Direct sequencing of large flavivirus PCR products for analysis of genome variation and molecular epidemiological investigations. , 1992, Journal of virological methods.

[5]  Takahiro Tomoyasu,et al.  Development of the immunomagnetic enrichment method selective for Vibrio parahaemolyticus serotype K and its application to food poisoning study , 1992, Applied and environmental microbiology.

[6]  M. Uhlén,et al.  Variations in the cytomegalovirus major immediate-early gene found by direct genomic sequencing , 1992, Journal of clinical microbiology.

[7]  P. E. Granum,et al.  Detection of Clostridium perfringens type A enterotoxin in faecal and food samples using immunomagnetic separation (IMS)-ELISA. , 1991, International journal of food microbiology.

[8]  E. Hornes,et al.  Pathogenic Escherichia coli found in food. , 1991, International journal of food microbiology.

[9]  M. Uhlén,et al.  General colorimetric method for DNA diagnostics allowing direct solid-phase genomic sequencing of the positive samples. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[10]  J. Albert,et al.  Few infected CD4+ T cells but a high proportion of replication-competent provirus copies in asymptomatic human immunodeficiency virus type 1 infection , 1991, Journal of virology.

[11]  M. Uhlén,et al.  Analysis of the V3 loop in neutralization-resistant human immunodeficiency virus type 1 variants by direct solid-phase DNA sequencing. , 1991, AIDS research and human retroviruses.

[12]  Robert L. Buchanan,et al.  Rapid isolation of Escherichia coli O157:H7 from enrichment cultures of foods using an immunomagnetic separation method , 1992 .

[13]  W. Hill,et al.  Loss of plasmids during enrichment for Escherichia coli , 1981, Applied and environmental microbiology.

[14]  N. Cross,et al.  A potent inhibitor of Taq polymerase copurifies with human genomic DNA. , 1988, Nucleic acids research.

[15]  E. Thorsby,et al.  Reliable isolation of human immunodeficiency virus from cultures of naturally infected CD4+ T cells. , 1989, Journal of virological methods.

[16]  M. Uhlén,et al.  Colorimetric detection of Plasmodium falciparum and direct sequencing of amplified gene fragments using a solid phase method. , 1992, Molecular and cellular probes.

[17]  M. Uhlén,et al.  Persistence of azidothymidine-resistant human immunodeficiency virus type 1 RNA genotypes in posttreatment sera , 1992, Journal of Virology.

[18]  A. Svennerholm,et al.  Molecular Pathogenesis of Gastrointestinal Infections , 1991, Federation of European Microbiological Societies Symposium Series.

[19]  D. Raoult,et al.  Diagnosis of Mediterranean spotted fever by indirect immunofluorescence of Rickettsia conorii in circulating endothelial cells isolated with monoclonal antibody-coated immunomagnetic beads. , 1992, The Journal of infectious diseases.

[20]  Ø. Olsvik,et al.  A nested PCR followed by magnetic separation of amplified fragments for detection of Escherichia coli Shiga-like toxin genes. , 1991, Molecular and cellular probes.

[21]  P. Muir,et al.  Rapid diagnosis of enterovirus infection by magnetic bead extraction and polymerase chain reaction detection of enterovirus RNA in clinical specimens , 1993, Journal of clinical microbiology.

[22]  M. Uhlén,et al.  Immunomagnetic recovery of Chlamydia trachomatis from urine with subsequent colorimetric DNA detection. , 1992, PCR methods and applications.

[23]  W. L. Payne,et al.  Isolation of Latin American epidemic strain of Vibrio cholerae 01 from US Gulf Coast , 1992, The Lancet.

[24]  D. Persing,et al.  Diagnostic molecular microbiology : principles and applications , 1993 .

[25]  G. Buffone Improved amplification of cytomegalovirus DNA from urine after purification of DNA with glass beads. , 1991, Clinical chemistry.

[26]  M. Vaudin,et al.  A rapid semi-automated microtiter plate method for analysis and sequencing by PCR from bacterial stocks. , 1989, Nucleic acids research.

[27]  M. Uhlén,et al.  Rapid colorimetric detection of in vitro amplified DNA sequences. , 1990, DNA and cell biology.

[28]  F. Vartdal,et al.  Rapid isolation of K88+ Escherichia coli by using immunomagnetic particles , 1988, Journal of clinical microbiology.

[29]  P. Gibbs,et al.  Separation and detection of salmonellae using immunomagnetic particles , 1991 .

[30]  Ø. Olsvik,et al.  Preparation and application of new monosized polymer particles , 1992 .

[31]  K. Krogfelt Bacterial adhesion: genetics, biogenesis, and role in pathogenesis of fimbrial adhesins of Escherichia coli. , 1991, Reviews of infectious diseases.

[32]  Y. Wasteson,et al.  Characterization of Escherichia coli strains isolated from pigs with edema disease. , 1992, Veterinary microbiology.

[33]  R. Beumer,et al.  Isolation of salmonellas by immunomagnetic separation. , 1992, The Journal of applied bacteriology.

[34]  J. Saunders,et al.  Rapid Immunocapture of Pseudomonas putida Cells from Lake Water by Using Bacterial Flagella , 1991, Applied and environmental microbiology.

[35]  Ø. Olsvik,et al.  Detection of Virulence Determinants in Enteric Escherichia coli Using Nucleic Acid Probes and Polymerase Chain Reaction , 1991 .

[36]  T E Michaelsen,et al.  Detection of pathogenic Yersinia enterocolitica in foods and water by immunomagnetic separation, nested polymerase chain reactions, and colorimetric detection of amplified DNA , 1993, Applied and environmental microbiology.

[37]  J. Jarp,et al.  A rapid assay for protein‐A in Staph, aureus strains, using immunomagnetic monosized polymer particles , 1988, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[38]  C Carrillo,et al.  The molecular epidemiology of cholera in Latin America. , 1993, The Journal of infectious diseases.

[39]  Ø. Olsvik,et al.  Detection of Escherichia coli heat-stable enterotoxin genes in pig stool specimens by an immobilized, colorimetric, nested polymerase chain reaction , 1991, Journal of clinical microbiology.

[40]  Max Sussman,et al.  New techniques in food and beverage microbiology. , 1993 .

[41]  M. Uhlén,et al.  Rapid detection and sequencing of specific in vitro amplified DNA sequences using solid phase methods. , 1990, Molecular and cellular probes.

[42]  E. Skjerve,et al.  Immunomagnetic separation of Salmonella from foods. , 1991, International journal of food microbiology.

[43]  A. Deelder,et al.  Magnetic bead antigen capture enzyme-linked immunoassay in microtitre trays for rapid detection of schistosomal circulating anodic antigen. , 1992, Journal of immunological methods.

[44]  D. Persing Polymerase chain reaction: trenches to benches , 1991, Journal of clinical microbiology.

[45]  R. Yolken,et al.  Removal of inhibitory substances from human fecal specimens for detection of group A rotaviruses by reverse transcriptase and polymerase chain reactions , 1990, Journal of clinical microbiology.

[46]  M. Uhlén,et al.  Magnetic separation of DNA , 1989, Nature.

[47]  M. Uhlén,et al.  Direct solid phase sequencing of genomic and plasmid DNA using magnetic beads as solid support. , 1989, Nucleic acids research.

[48]  Y. Wasteson,et al.  Immunomagnetic separation and DNA hybridization for detection of enterotoxigenic Escherichia coli in a piglet model , 1991, Journal of clinical microbiology.

[49]  G. Schoolnik,et al.  Multi‐gene amplification: simultaneous detection of three virulence genes in diarrhoeal stool , 1989, Molecular microbiology.

[50]  K. A. Fleming,et al.  FALSE-POSITIVE RESULTS AND THE POLYMERASE CHAIN REACTION , 1988, The Lancet.

[51]  C. Lattuada,et al.  Isolation of Escherichia coli 0157:H7 Using 0157 Specific Antibody Coated Magnetic Beads. , 1992, Journal of food protection.

[52]  D. Relman,et al.  Identification of the uncultured bacillus of Whipple's disease. , 1992, The New England journal of medicine.

[53]  A. Lindberg,et al.  Detection of Shigella dysenteriae type 1 and Shigella flexneri in feces by immunomagnetic isolation and polymerase chain reaction , 1992, Journal of clinical microbiology.

[54]  M. Uhlén,et al.  Affinity Separation of Nucleic Acids on Monosized Magnetic Beads , 1993 .

[55]  J. Verhoef,et al.  The magnetic immuno polymerase chain reaction assay for direct detection of salmonellae in fecal samples , 1992, Journal of clinical microbiology.

[56]  L. Mayer Use of plasmid profiles in epidemiologic surveillance of disease outbreaks and in tracing the transmission of antibiotic resistance , 1988, Clinical Microbiology Reviews.

[57]  T. Popović,et al.  Use of polymerase chain reaction for detection of toxigenic Vibrio cholerae O1 strains from the Latin American cholera epidemic , 1992, Journal of clinical microbiology.

[58]  H. Ushijima,et al.  Removal of HIV antigens and HIV-infected cells in vitro using immunomagnetic beads. , 1990, Journal of virological methods.

[59]  E. Hornes,et al.  Identification of a double-stranded RNA virus by using polymerase chain reaction and magnetic separation of the synthesized DNA segments , 1990, Journal of clinical microbiology.

[60]  J. Saunders,et al.  Detection of genetically engineered traits among bacteria in the environment. , 1990, Trends in biotechnology.

[61]  M. Uhlén,et al.  Immunomagnetic purification to facilitate DNA diagnosis of Plasmodium falciparum , 1993, Journal of clinical microbiology.

[62]  D. Talkington,et al.  Detection of streptococcal pyrogenic exotoxin genes by a nested polymerase chain reaction. , 1993, Molecular and cellular probes.

[63]  J. Luk,et al.  Rapid and sensitive detection of Salmonella (O:6,7) by immunomagnetic monoclonal antibody-based assays. , 1991, Journal of immunological methods.

[64]  Barry Goldstein,et al.  Optimization of the controlled separation of biologically active diagnostic magnetic probes , 1990 .

[65]  J. Verhoef,et al.  Monoclonal antibodies that detect live salmonellae , 1992, Applied and environmental microbiology.

[66]  M. Uhlén,et al.  Dynamic changes in HIV‐1 quasispecies from azidothymidine (AZT)‐treated patients , 1992, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[67]  P. Parham,et al.  Direct binding of influenza peptides to class I HLA molecules , 1989, Nature.