Skip directly to site content Skip directly to page options Skip directly to A-Z link Skip directly to A-Z link Skip directly to A-Z link
Volume 7, Number 2—April 2001
THEME ISSUE
4th Decennial International Conference on Nosocomial and Healthcare-Associated Infections
State of the Art

Molecular Approaches to Diagnosing and Managing Infectious Diseases: Practicality and Costs

Michael A. PfallerComments to Author 
Author affiliation: University of Iowa College of Medicine, Iowa City, Iowa, USA

Main Article

Table 4

Genotypic methods for epidemiologic typing of microorganismsa,b

Method Examples Comments
Plasmid analysis Staphylococci Enterobacteriaceae Plasmids may be digested with restriction endonucleases
Only useful when organisms carry plasmids
Restriction endonuclease analysis of chromosomal DNA with conventional electrophoresis Enterococci
Staphylococcus aureus
Clostridium difficile
Candida spp. Large number of bands Difficult to interpret
Not amenable to computer analysis
PFGE Enterobacteriaceae
Staphylococci
Enterococci
Candida spp. Fewer bands
Amenable to computer analysis
Very broad application.
Genome restriction fragment length polymorphism analysis: ribotyping, insertion sequence probe fingerprinting Enterobacteriaceae
Staphylococci
Pseudomonas aeruginosa
Mycobacterium tuberculosis
Candida spp. Fewer bands
Computer analysis
Sequence-based profiles
Automated
PCR-based methods: repetitive elements PCR spacer typing, selective amplification of genome restriction fragments, multilocus allelic sequence-based typing Enterobacteriaceae
Acinetobacter spp.
Staphylococci
M. tuberculosis
HCV Crude extracts and small amounts of DNA may suffice
Library probe genotypic hybridization schemes: multilocus probe dot-blot patterns, high-density oligonucleotide patterns Burkholderia cepacia
S. aureus
M. tuberculosis Unambiguous yes-no result
Less discrimination than other methods
Couple with DNA chip technology

aThe table contains examples of available methods and applications and is not intended to be all-inclusive.
bAdapted from Pfaller (2).

Main Article

References
  1. Cormican  MG, Pfaller  MA. Molecular pathology of infectious diseases. In: Henry JB, editor. Clinical diagnosis and management by laboratory methods. 19th ed. Philadelphia: W.B. Saunders Company; 1996:1390-9.
  2. Pfaller  MA. Diagnosis and management of infectious diseases: Molecular methods for the new millennium. Clinical Laboratory News. 2000;26:103.
  3. Kant  JA. Molecular diagnostics: Reimbursement and other selected financial issues. Diagn Mol Pathol. 1995;4:7981. DOIPubMed
  4. Fredricks  DN, Relman  DA. Sequence-based identification of microbial pathogens: A reconsideration of Koch's postulates. Clin Microbiol Rev. 1996;9:1833.PubMed
  5. Fredricks  DN, Relman  DA. Application of polymerase chain reaction to the diagnosis of infectious disease. Clin Infect Dis. 1999;29:47588. DOIPubMed
  6. Tang  YW, Persing  DH. Molecular detection and identification of microorganisms. In: Murray PR, Baron EJ, Pfaller MA, Tenover FC, Yolken RH, editors. Manual of clinical microbiology. 7th ed. Washington: American Society for Microbiology; 1999:215-44.
  7. Woods  GL. Molecular techniques in mycobacterial detection. Arch Pathol Lab Med. 2001;125:1226.PubMed
  8. Bergeron  MG, Ouellette  M. Preventing antibiotic resistance using rapid DNA-based diagnostic tests. Infect Control Hosp Epidemiol. 1998;19:5604. DOIPubMed
  9. Cockerill  FR III. Genetic methods for assessing antimicrobial resistance. Antimicrob Agents Chemother. 1999;43:199212.PubMed
  10. Hacek  DM, Suriano  T, Noskin  GA, Kruszynski  J, Reisberg  B, Peterson  LR. Medical and economic benefit of a comprehensive infection control program that includes routine determination of microbial clonality. Am J Clin Pathol. 1999;111:64754.PubMed
  11. Ross  JS. Financial determinants of outcomes in molecular testing. Arch Pathol Lab Med. 1999;123:10715.PubMed
  12. Pfaller  MA. Molecular epidemiology in the care of patients. Arch Pathol Lab Med. 1999;123:100710.PubMed
  13. Nolte  FS. Impact of viral load testing on patient care. Arch Pathol Lab Med. 1999;123:10114.PubMed
  14. Anthony  RM, Brown  TJ, French  GL. Rapid diagnosis of bacteremia by universal amplification of 23S ribosomal DNA followed by hybridization to an oligonucleotide array. J Clin Microbiol. 2000;38:7818.PubMed
  15. Marshall  SA, Wilke  WW, Pfaller  MA, Jones  RN. Staphylococcus aureus and coagulase-negative staphylococci from blood stream infections: Frequency of occurrence, antimicrobial susceptibility, and molecular (mec A) characterization of oxacillin resistance in the SCOPE Program. Diagn Microbiol Infect Dis. 1998;30:20514. DOIPubMed
  16. Hussain  Z, Stoakes  L, Massey  V, Diagre  D, Fitzgerald  V, El Sayed  S, Correlation of oxacillin MIC with mec A gene carriage in coagulase-negative staphylococci. J Clin Microbiol. 2000;38:7524.PubMed
  17. Hecht  FM, Grant  RM, Petropoulos  CJ, Dillon  B, Chesney  MA, Tian  H, Sexual transmission of an HIV-1 variant resistant to multiple reverse-transcriptase and protease inhibitors. N Engl J Med. 1998;339:30711. DOIPubMed
  18. Stuyver  L, Van Geyt  C, de Gendt  S, Van Reybroeck  G, Zoulin  F, Leroux-Rods  G, Line probe assay for monitoring drug resistance in hepatitis B virus-infected patients during antiviral therapy. J Clin Microbiol. 2000;38:7027.PubMed
  19. Courvalin  P. Genotypic approach to the study of bacterial resistance to antibiotics. Antimicrob Agents Chemother. 1991;35:101923.PubMed
  20. Arbeit  RD. Laboratory procedures for epidemiologic analysis of microorganisms. In: Murray PR, Baron EJ, Pfaller MA, Tenover FC, Yolken RH, editors. Manual of clinical microbiology. 7th ed. Washington: American Society for Microbiology; 1999:116-37.
  21. Pfaller  MA, Herwaldt  LA. The clinical microbiology laboratory and infection control: Emerging pathogens, antimicrobial resistance, and new technology. Clin Infect Dis. 1997;25:85870. DOIPubMed
  22. Ferreira-Gonzalez  A, Garrett  CG. Pitfalls in establishing a molecular diagnostic laboratory. Hum Pathol. 1996;27:43740. DOIPubMed
  23. Back  NA, Linnemann  CC, Pfaller  MA, Staneck  JL, Morthland  V. Recurrent epidemics caused by a single strain of erythromycin-resistant Staphylococcus aureus: The importance of molecular epidemiology. JAMA. 1993;270:132933. DOIPubMed

Main Article

Page created: May 10, 2011
Page updated: May 10, 2011
Page reviewed: May 10, 2011
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.
file_external