Microbiology

Lung/Mycobacterial Research Advances


Introduction
The following is a list of some of the accomplishments of the Department of Microbiology over the past 20 years. Since 1985, the department has had more than 300 scientific publications, most related to diagnosis, treatment, new drugs, and the genetics of drug resistance in mycobacteria. A summary of the major accomplishments and some related publications are
listed below.

Research Summary

I. Nomenclature/Taxonomy (the naming of species and determination of their relationships to other species)

1.The discovery, description, and naming (in collaboration with other national and international collaborators) of one new species of Nocardia and four new species and two biovariants (subspecies) of mycobacteria: (1-4)
Nocardia pseudobrasiliensis
Mycobacterium mucogenicum
Mycobacterium goodii
Mycobacterium wolinskyi
Mycobacterium immunogenum
Mycobacterium fortuitum third biovariant sorbitol positive
Mycobacterium fortuitum third biovariant sorbitol negative
2.Description of nontypable Haemophilus influenzae biotype IV as a neonatal and genital tract pathogen.(5)
3.Description of first isolates of tetracycline and erythromycin resistant Branhamella (Moraxella) catarrhalis. (33)

II. Molecular typing/DNA Fingerprinting (Genetic methods to study hospital outbreaks or multiple cultures from the same patient to see if the cultures are identical or different. Also can be used for rapid species identification by molecular methods.)

  1. Development of molecular typing methods for comparision of different strains of rapidly growing mycobacteria and M. avium complex, including the techniques of pulsed field gel electrophoresis and random amplified polymorphic DNA polymerase chain reaction
    (PCR). (For examples of its use, see references 6-10.)
  2. Development of PCR restriction fragment length polymorphism analysis (PRA) for rapid species identification of Nocardia and rapidly growing mycobacteria. (This method currently used in our lab for identification to species of all cultures of Nocardia and both rapidly growing and slowly growing Mycobacteria.) We are one of the first laboratories to routinely utilize a nonsequencing PCR method for such identification. (For examples of its use, see references 11-13.)

III. Drug Resistance Mechanisms

  1. Description of the genetic mutation in the 16S ribosomal RNA gene responsible for resistance to clarithromycin in M. chelonae, M. abscessus, M. avium, and M. intracelluare. (14-15)
  2. 2.Description of the genetic mutation in the 23S ribosomal RNA gene responsible for the development of resistance to amikacin in M. chelonae, M. abscessus, M. peregrinum, and M. fortuitum. (Amikacin is an important injectable medicine for each of these species.) (16)
  3. 3.Characterization of the penicillinase that confers resistance to penicillin and related drugs (called BRO ß-lactamase) in Branhamella catarrhalis (a common lung pathogen in patients with chronic lung disease), and its earliest appearance among clinical strains in the United States. (17)
  4. 4.Characterization of ß-lactamases (enzymes that inactivate penicillins and related compounds) in Mycobacterium tuberculosis, nocardia, and rapidly growing mycobacteria. (18-20)

IV. Susceptibility Testing (Determination of whether a drug is likely to be useful and effective when given to a patient.)

  1. Recognition of the activity (potential clinical usefulness) of clarithromycin and other newer macrolides against rapidly growing and some slowly growing nontuberculous mycobacterial species, including against M. chelonae, M. abscessus, M. fortuitum, M. kansasii, M. marinum, and M. haemophilum. (This drug is the antibiotic of choice for many of these species.) (21-22)
  2. Recognition of the activity of the new compound linezolid against Nocardia. (34)
  3. Recognition of the activity of linezolid against rapidly growing mycobacteria, especially against M. fortuitum and M. chelonae. (35)
  4. Recognition of the activity of the tetracycline related compounds, the glycylcyclines, against the rapidly growing mycobacteria. (23)
  5. .Development (along with other committee members) of the first Clinical Laboratory Standards Institute, CLSI, formerly National Committeee for Clinical Laboratory Standards (NCCLS) standard for susceptibility testing of Nocardia and nontuberculous mycobacteria. NCCLS M24-A, 2003

V. Diagnosis and Treatment of Disease

  1. Preparing the first (1990) and the second (1997) American Thoracic Society Statements on Diagnosis and Treatment of Nontuberculous Mycobacteria. These are the major United States guidelines regarding diagnosis and treatment of species such as M. avium complex (MAC), M. kansasii, and the rapidly growing mycobactera. (24)
  2. First and only description of treatment regimens for rifampin resistant M. kansasii. (25)
  3. First to recognize that multiple strains (different organisms), not just one, are rountinely present in M. avium complex infection in nodular bronchiectasis. (26)
  4. Development of three times weekly treatment regimen currently recommended for treatment of pulmonary MAC and recognition that it was as effective as daily therapy. (27-28, 36)
  5. Development of the standard 3-drug macrolide containing regimens for treatment of pulmonary MAC. (29)
  6. The first treatment trials in the United States to show the efficacy of clarithromycin (above) for the treatment of pulmonary MAC. (30-31)
  7. Performance of the first and only clinical trial of clarithromycin for treatment of Mycobacterium chelonae, and establishment of clarithromycin (biaxin) as the drug of choice for this infection. (32)

RESEARCH SUMMARY REFERENCES

  1. Wallace RJ Jr, Brown BA, Silcox VA, Tsukamura M, Nash DR, Steele LC, Steingrube VA, Smith J, Sumter G, Zhang Y, Blacklock Z: Clinical disease, drug susceptibility, and biochemical patterns of the unnamed third biovariant complex of Mycobacterium fortuitum. J. Infect. Dis. 163:598-603, 1991.
  2. Springer B, Böttger EC, Kirschner P, Wallace RJ Jr: Phylogeny of the Mycobacterium chelonae-like organism based on partial sequencing of the 16S rRNA gene and proposal of Mycobacterium mucogenicum sp. nov. Intern. J. Syst. Bacteriol. 45:262-267, 1995.
  3. Ruimy R, Riegel P, Carlotti A, Boiron P, Bernardin G, Monteil H, Wallace RJ Jr, Christen R: Nocardia pseudobrasiliensis sp. nov., a new species of Nocardia which groups bacterial strains previously identified as Nocardia brasiliensis and associated with invasive diseases. Intern. J. Syst. Bacteriol. 46:259-264, 1996.
  4. Brown BA, Springer B, Steingrube VA, Wilson RW, Pfyffer GE, Garcia MJ, Menendez MC, Rodriguez-Salgado B, Jost KC Jr., Chiu SH, Onyi GO, Böttger EC, Wallace RJ Jr: Mycobacterium wolinskyi sp. nov. and Mycobacterium goodii sp. nov., two new rapidly growing species related to Mycobacterium smegmatis and associated with human wound infections: a cooperative study from the International Working Group on Mycobacterial Taxonomy. Intern. J. Syst. Bacteriol. 49:1493-1511, 1999.
  5. Quentin R, Goudeau A, Wallace RJ Jr, Smith AL, Selander RK, Musser JM: Urogenital, maternal, and neonatal isolates of Haemophilus influenzae: Identification of unusually virulent serologically nontypable clone families and strong evidence for a new Haemophilus species. J. Clin. Microbiol. 136:1203-1209, 1990.
  6. Hector JSR, Pang Y, Mazurek GH, Zhang Y, Brown BA, Wallace RJ Jr: Large restriction fragment patterns of genomic Mycobacterium fortuitum DNA as strain-specific markers and their use in epidemiologic investigation of four nosocomial outbreaks. J. Clin. Microbiol. 30:1250-1255, 1992.
  7. Mazurek GH, Hartman S, Zhang Y-S, Brown BA, Hector JSR, Murphy D, Wallace RJ Jr: Large DNA restriction fragment polymorphism in the Mycobacterium avium-M. intracellulare complex: A potential epidemiologic tool. J. Clin. Microbiol. 31:390-394, 1993.
  8. Maloney S, Welbel S, Daves B, Adams K, Becker S, Bland L, Arduino M, Wallace RJ Jr., Zhang Y, Buck G, Risch P, Jarvis W: Mycobacterium abscessus pseudoinfection traced to an automated endoscope washer: Utility of epidemiologic and laboratory investigation. J. Infect. Dis. 169:1166-1169, 1994.
  9. Zhang Y, Rajagopalan M, Brown BA, Wallace RJ Jr: Randomly amplified polymorphic DNA PCR for comparison of Mycobacterium abscessus strains from nosocomial outbreaks. J. Clin. Microbiol. 35:3132-3139, 1997.
  10. Wallace RJ Jr, Brown BA, Griffith DE: Nosocomial outbreaks/pseudo-outbreaks caused by nontuberculous mycobacteria. Annu. Rev. Microbiol. 52:453-490, 1998.
  11. Steingrube VA, Gibson JL, Brown BA, Zhang Y, Wilson RW, Rajagopalan M, Wallace RJ Jr: PCR amplification and restriction endonuclease analysis of a 65-kilodalton heat shock protein gene sequence for taxonomic separation of rapidly growing mycobacteria. J. Clin. Microbiol. 33:149-153, 1995.
  12. .Steingrube, VA, Wilson RW, Brown BA, Jost KC Jr., Blacklock Z, Gibson JL, Wallace RJ Jr: Rapid identification of clinically significant species and taxa of aerobic actinomycetes, including Actinomadura, Gordona, Nocardia, Rhodococcus, Streptomyces, and Tsukamurella isolates, by DNA amplification and restriction endonuclease analysis. J. Clin. Microbiol. 35:817-822, 1997.
  13. Wilson RW, Steingrube VA, Brown BA, Wallace RJ Jr: Clinical application of PCR-restriction enzyme pattern analysis for rapid identification of aerobic actinomycete isolates. J. Clin. Microbiol. 36:148-152, 1998.
  14. Wallace RJ Jr., Meier A, Brown BA, Zhang Y, Sander P, Onyi GO, Böttger EC: Genetic basis for clarithromycin resistance among isolates of Mycobacterium chelonae and Mycobacterium abscessus. Antimicrob. Agents Chemother. 40:1676-1681, 1996.
  15. Meier A, Heifets L, Wallace RJ Jr., Zhang Y, Brown BA, Sander P, Böttger EC: Molecular mechanisms of clarithromycin resistance in Mycobacterium avium: observation of multiple 23S rDNA mutations in a clonal population. J. Infect. Dis. 174:354-360, 1996.
  16. Prammananan T, Sander P, Brown BA, Frischkorn K, Onyi GO, Zhang Y, Böttger EC, Wallace RJ Jr: A single 16S ribosomal RNA substitution is responsible for resistance to amikacin and other 2-deoxystreptamine aminoglycosides in Mycobacterium abscessus and Mycobacterium chelonae. J. Infect Dis. 177:1573-1581, 1998.
  17. Wallace RJ Jr, Steingrube VA, Nash DR, Hollis D, Flanagan C, Brown BA, Labidi A, Weaver R: BRO ß-Lactamases of Branhamella catarrhalis and subgenus Moraxella, including evidence for chromosomal ß-lactamase transfer by conjugation in B. catarrhalis, M. nonliquefaciens, and M. lacunata. Antimicrob. Agents Chemother. 33:1845-1854, 1989.
  18. Zhang Y, Steingrube VA, Wallace RJ Jr: ß-lactamase Inhibitors and the inducibility of the beta-lactamase of Mycobacterium tuberculosis. Am Rev. Respir. Dis. 145:657-660, 1992.
  19. Zhang Y, Wallace RJ Jr, Steingrube VA, Brown BA, Nash DR, Silcox VA, Tsukamura M: Isoelectric focusing patterns of ß-lactamases in the rapidly growing Mycobacteria. Tuberc. Lung Dis. 73:337-344, 1992.
  20. Steingrube VA, Wallace RJ Jr, Brown BA, Zhang Y, Steele LC, Young G, Nash DR: Partial characterization of Nocardia farcinica ß-lactamases. Antimicrob. Agents Chemother. 37:1850-1855, 1993.
  21. Brown BA, Wallace RJ Jr, Onyi G, DeRosas V, Wallace RJ III: Activities of four macrolides including clarithromycin against Mycobacterium fortuitum, Mycobacterium chelonae, and Mycobacterium chelonae-like organisms. Antimicrob. Agents Chemother. 36:180-184, 1992.
  22. Brown BA, Wallace RJ Jr, Onyi G: Activities of clarithromycin against eight slowly growing species of nontuberculous mycobacteria, determined by using a broth microdilution MIC system. Antimicrob. Agents Chemother. 36:1987-1990, 1992.
  23. Brown BA, Wallace RJ Jr., Onyi GO: Activities of the glycylcyclines N, N- dimethylglycylamido-minocycline and N, N-dimethylglycylamido-6-demethyl-6- deoxytetracycline against Nocardia spp. and tetracycline-resistant isolates of rapidly growing mycobacteria. Antimicrob. Agents Chemother. 40:874-878, 1996.
  24. Wallace RJ Jr., Cook JL, Glassroth J, Griffith DE, Olivier KN, Gordin F: Diagnosis and treatment of disease caused by nontuberculous mycobacteria. American Thoracic Society Statement. Am. J. Resp. Crit. Care Med. 156:S1-S25, 1997.
  25. Wallace RJ Jr, Dunbar D, Brown BA, Onyi G, Dunlap R, Ahn CH, Murphy D: Rifampin-resistant Mycobacterium kansasii. Clin. Infect. Dis. 18:736-743, 1994.
  26. Wallace RJ Jr, Zhang Y, Brown BA, Dawson D, Murphy DT, Wilson R, Griffith DE: Polyclonal Mycobacterium avium complex infections in patients with nodular bronchiectasis. Am. J. Respir. Crit. Care Med. 158:1235-1244, 1998.
  27. Griffith DE, Brown BA, Murphy DT, Girard WM, Couch L, Wallace RJ Jr: Initial (6-month) results of three-times-weekly azithromycin in treatment regimens for Mycobacterium avium complex lung disease in human immunodeficiency virus - negative patients. J. Infect. Dis. 178:121-126, 1998.
  28. Griffith DE, Brown BA, Cegielski P, Murphy DT, Wallace RJ Jr: Early results (at 6 months) with intermittent clarithromycin-including regimens for lung disease due to Mycobacterium avium complex. Clin. Infect. Dis. 30:288-292, 2000.
  29. Griffith DE, Wallace RJ Jr: Treatment of pulmonary Mycobacterium avium complex lung disease in non-acuired immunodeficiency syndrome (AIDS) patients in the era of the newer macrolides and rifabutin. Am. J. Med. 102:22-27, 1997.
  30. Wallace RJ Jr, Brown BA, Griffith DE, Girard WM, Murphy DT, Onyi GO, Steingrube VA, Mazurek GH: Initial clarithromycin monotherapy for Mycobacterium avium-intracellulare complex lung disease. Amer. Rev. Resp. Dis. 149:1335-1341, 1994.
  31. Wallace RJ Jr, Brown BA, Griffith DE, Girard WM, Murphy DT: Clarithromycin regimens for pulmonary Mycobacterium avium complex - the first 50 patients. Am. J. Resp. Crit. Care Med. 153:1766-1772, 1996.
  32. .Wallace RJ Jr, Tanner D, Brennan PJ, Brown BA: Clinical trial of clarithromycin for cutaneous (disseminated) infection due to Mycobacterium chelonae. Ann. Intern. Med. 119:482-486, 1993.
  33. Brown BA, Wallace RJ Jr, Flanagan CW, Wilson RW, Luman JL, Redditt SD: Tetracycline and erythromycin resistance among clinical isolates of Branhamella catarrhalis. Antimicrob. Agents Chemother. 33:1631-1633, 1989.
  34. Brown BA, Ward SC, Crist CJ, Mann LB, Wilson RW, Wallace RJ Jr: In vitro activities of linezolid against multiple Nocardia species. Antimicrob. Agents Chemother. 45:1295-1297, 2001.
  35. Wallace RJ Jr, Brown BA, Ward SC, Crist CJ, Mann LB, Wilson RW: Activities of linezolid against rapidly growing mycobacteria. Antimicrob. Agents Chemother. 45:764-767, 2001.
  36. Griffith DE, Brown BA, Cegielski P, Murphy DT, Wallace RJ Jr: Early results (at 6 Months) with intermittent clarithromycin-including regimens for lung disease due to Mycobacterium avium complex. Clin. Inf. Dis. 30:288-92, 2000.

Last Update: October 10, 2006