Analysis of Bioaerosol Samples

Patricia Cruz; Mark P. Buttner

doi:10.1128/9781555815882.ch75

Chapter 75 : Analysis of Bioaerosol Samples

Authors: Patricia Cruz, Mark P. Buttner

Content Type: Reference

Publication Year: 2007

Category: Environmental Microbiology

Book DOI: 10.1128/9781555815882

Chapter DOI: 10.1128/9781555815882.ch75

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Preview this chapter:

Analysis of Bioaerosol Samples, Page 1 of 2

< Previous page | Next page > /docserver/preview/fulltext/10.1128/9781555815882/9781555813796_Chap75-1.gif /docserver/preview/fulltext/10.1128/9781555815882/9781555813796_Chap75-2.gif

Abstract:

Respiratory exposure to certain pathogenic or toxigenic microorganisms and/or elevated concentrations of environmental organisms could result in health effects, such as allergic reactions, irritant responses, toxicosis, and respiratory illness. This chapter presents an overview of available methods for the analysis of bioaerosols. In addition, the potential use of enhanced monitoring of bioaerosols with PCR, biochemical, and immunological assays is discussed. Several conditions, such as pH, temperature, water activity, nutrients, antibiotics, light, and aeration, can be manipulated to favor the growth of a selected group of organisms. In contrast with culture techniques, microscopic analysis allows enumeration of both culturable and nonculturable microorganisms. However, identification of microorganisms to the species level is usually not possible without the aid of a taxon-specific technique, such as immunospecific fluorescence staining. Flow cytometry offers an alternative to microscopic enumeration of total cells. In a different application of flow cytometry, researchers coupled sandwich immunoassay utilizing microsphere beads with flow cytometry for the detection of biothreat agents. Fluorescence immunoassay consists of staining samples with a fluorescently labeled antibody that binds specifically to the antigens on the surfaces of the target organisms and enumeration by epifluorescence microscopy. Hybrid technologies are being developed that combine more than one analysis method to maximize the accuracy of the results obtained. One such system utilizes immunoassay, flow cytometry, and PCR analysis in various combinations for the continuous monitoring of airborne biological threat agents.

Citation: Cruz P, Buttner M. 2007. Analysis of Bioaerosol Samples, p 952-960. In Hurst C, Crawford R, Garland J, Lipson D, Mills A, Stetzenbach L (ed), Manual of Environmental Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815882.ch75

Highlighted Text: Show | Hide

Full text loading...

Figures

Analysis of Bioaerosol Samples

[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555815882.ch75-1,webId=/content/author/10.1128/9781555815882.ch75-1,properties={foaf_givenname=Patricia, foaf_name=Patricia Cruz, foaf_surname=Cruz}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555815882.ch75-2,webId=/content/author/10.1128/9781555815882.ch75-2,properties={foaf_givenname=Mark P., foaf_name=Mark P. Buttner, foaf_surname=Buttner}]]

/content/10.1128/9781555815882.ch75.ch75fig01

FIGURE 1

Diagrams illustrating competitive PCR (cPCR) and QPCR. (A) In cPCR, the target template and the internal control DNA compete for the oligonucleotide primers during the PCR assay. Post-PCR manipulations are required to separate the target and competitor products and to determine the ratio between them (i.e., final product concentrations). (B) In QPCR, a fluorescent probe anneals to the target DNA between the specific primer binding sites. As new DNA is synthesized, the probe is cleaved by the Taq polymerase, causing it to fluoresce; the amount of fluorescence is used to measure the initial target DNA concentration.

10.1128/9781555815882/f0956-01_thmb.gif

10.1128/9781555815882/f0956-01.gif

FIGURE 1

References

/content/book/10.1128/9781555815882.ch75

1. Alvarez, A. J.,, M. P. Buttner,, G. A. Toranzos,, E. A. Dvorsky,, A. Toro,, T. B. Heikes,, L. E. Mertikas, and, L. D. Stetzenbach. 1994. The use of solid-phase polymerase chain reaction for the enhanced detection of airborne microorganisms. Appl. Environ. Microbiol. 60: 374– 376.

2. Alvarez, A. J.,, M. P. Buttner, and, L. D. Stetzenbach. 1995. PCR for bioaerosol monitoring: sensitivity and environmental interference. Appl. Environ. Microbiol. 61: 3639– 3644.

3. Andersen, A. A. 1958. New sampler for the collection, sizing, and enumeration of viable airborne particles. J. Bacteriol. 76: 471– 484.

4. Andreotti, P. E.,, G. V. Ludwig,, A. H. Peruski,, J. J. Tuite,, S. S. Morse, and, L. F. Peruski, Jr. 2003. Immunoassay of infectious agents. BioTechniques 35: 850– 859.

5. Banks, J. G.,, R. G. Board,, J. Carger, and, A. D. Dodge. 1985. The cytotoxic and photodynamic inactivation of microorganisms by rose bengal. J. Appl. Bacteriol. 58: 392– 400.

6. Bassler, H. A.,, S. J. A. Flood,, K. J. Livak,, J. Marmaro,, R. Knorr, and, C. A. Batt. 1995. Use of a fluorogenic probe in a PCR–based assay for the detection of Listeria monocytogenes. Appl. Environ. Microbiol. 61: 3724– 3728.

7. Beuchat, L. R. 1992. Media for detecting and enumerating yeasts and moulds. Int. J. Food Microbiol. 17: 145– 158.

8. Burge, H. A. 1995. Bioaerosol investigations, p. 1–23. In H. A. Burge (ed.), Bioaerosols. CRC Press, Boca Raton, Fla.

9. Burge, H. A.,, and W. R. Solomon. 1987. Sampling and analysis of biological aerosols. Atmos. Environ. 21: 451– 456.

10. Burge, H. P.,, W. R. Solomon, and, J. R. Boise. 1977. Comparative merits of eight popular media in aerometric studies of fungi. J. Allergy Clin. Immunol. 60: 199– 203.

11. Buttner, M. P.,, P. Cruz,, L. D. Stetzenbach,, A. K. Klima-Comba,, V. L. Stevens, and, T. D. Cronin. 2004. Determination of the efficacy of two building decontamination strategies by surface sampling with culture and quantitative PCR analysis. Appl. Environ. Microbiol. 70: 4740– 4747.

12. Buttner, M. P.,, P. Cruz,, L. D. Stetzenbach,, A. K. Klima-Comba,, V. L. Stevens, and, P. A. Emanuel. 2004. Evaluation of the Biological Sampling Kit (BiSKit) for large-area surface sampling. Appl. Environ. Microbiol. 70: 7040– 7045.

13. Buttner, M. P.,, P. Cruz-Perez, and, L. D. Stetzenbach. 2001. Enhanced detection of surface-associated bacteria in indoor environments by quantitative PCR. Appl. Environ. Microbiol. 67: 2564– 2570.

14. Buttner, M. P.,, and L. D. Stetzenbach. 1991. Evaluation of four aerobiological sampling methods for the retrieval of aerosolized Pseudomonas syringae. Appl. Environ. Microbiol. 57: 1268– 1270.

15. Buttner, M. P.,, and L. D. Stetzenbach. 1993. Monitoring airborne fungal spores in an experimental indoor environment to evaluate sampling methods and the effects of human activity on air sampling. Appl. Environ. Microbiol. 59: 219– 226.

16. Castellan, R. M.,, S. A. Olenchock,, K. B. Kinsley, and, J. L. Hankinson. 1987. Inhaled endotoxin and decreased spirometric values: an exposure-response relation for cotton dust. New Engl. J. Med. 317: 605– 610.

17. Chamberlain, J. S.,, R. A. Gibbs,, J. E. Ranier,, P. N. Nguyen, and, C. T. Cashey. 1988. Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification. Nucleic Acids Res. 16: 11141– 11156.

18. Chang, C. W.,, Y. H. Hwang,, S. A. Grinshpun,, J. M. Macher, and, K. Willeke. 1994. Evaluation of counting error due to colony masking in bioaerosol sampling. Appl. Environ. Microbiol. 60: 3732– 3738.

19. Chapman, M. D. 1995. Analytical methods: immunoas-says, p. 235–248. In H. A. Burge (ed.), Bioaerosols. Lewis Publishers, Boca Raton, Fla.

20. Chung, Y.-J.,, B. B. Jarvis,, H. Tak, and, J. J. Pestka. 2003. Immunochemical assay for satratoxin G and other macro-cyclic trichothecenes associated with indoor air contamination by Stachybotrys chartarum. Toxicol. Mech. Methods 13: 247– 252.

21. Croft, W. S.,, B. B. Jarvis, and, C. S. Yatawara. 1986. Airborne outbreak of trichothecene toxicosis. Atmos. Environ. 20: 549– 552.

22. Crook, B.,, S. Higgins, and, J. Lacey. 1987. Methods for sampling microorganisms at solid waste disposal sites, p. 791–797. In D. R. Houghton,, R. N. Smith, and, H. O. W. Eggins (ed.), Biodeterioration, vol. 7. Proceedings of the 7th International Biodeterioration Symposium. Elsevier, London, United Kingdom.

23. Cruz, P.,, and L. D. Stetzenbach. 2004. Specific detection of fungi associated with sick building syndrome when using quantitative polymerase chain reaction, p. 437–449. In D. C. Straus (ed.), Advances in Applied Microbiology, vol. 55. Sick Building Syndrome. Elsevier Academic Press, San Diego, Calif.

24. Cruz–Perez, P.,, M. P. Buttner, and, L. D. Stetzenbach. 2001. Detection and quantitation of Aspergillus fumigatus in pure culture using polymerase chain reaction. Mol. Cell. Probes 15: 81– 88.

25. Cruz–Perez, P.,, M. P. Buttner, and, L. D. Stetzenbach. 2001. Specific detection of Stachybotrys chartarum in pure culture using quantitative polymerase chain reaction. Mol. Cell. Probes 15: 129– 138.

26. de Aguirre, L.,, S. F. Hurst,, J. S. Choi,, J. H. Shin,, H. P. Hinrikson, and, C. J. Morrison. 2004. Rapid differentiation of Aspergillus species from other medically important opportunistic molds and yeasts by PCR-enzyme immuno-assay. J. Clin. Microbiol. 42: 3495– 3504.

27. Dean, T. R.,, B. Roop,, D. Betancourt, and, M. Y. Menetrez. 2005. A simple multiplex polymerase chain reaction assay for the identification of four environmentally relevant fungal contaminants. J. Microbiol Methods 61: 9– 16.

28. DeBlay, F.,, M. D. Chapman, and, T. A. Platts-Mills. 1991. Airborne cat allergen Feld I. Environmental control with the cat in situ. Am. Rev. Respir. Dis. 143: 1334– 1339.

29. Desjardin, L. E.,, Y. Chen,, M. D. Perkins,, L. Teixeira,, M. D. Cave, and, K. D. Eisenach. 1998. Comparison of the ABI 7700 System (TaqMan) and competitive PCR for quantification of IS 6110 DNA in sputum during treatment of tuberculosis. J. Clin. Microbiol. 36: 1964– 1968.

30. Douwes, J.,, G. Doekes,, R. Montijn,, D. Heederik, and, B. Brunekreef. 1996. Measurement of β(1→3)-glucans in occupational and home environments with an inhibition enzyme immunoassay. Appl. Environ. Microbiol. 62: 3176– 3182.

31. Douwes, J.,, P. Thorne,, N. Pearce, and, D. Heederik. 2003. Bioaerosol health effects and exposure assessment: progress and prospects. Ann. Occup. Hyg. 47: 187– 200.

32. Eduard, W.,, and D. Heederik. 1998. Methods for quantitative assessment of airborne levels of noninfectious microorganisms in highly contaminated work environments. Am. Ind. Hyg. Assoc. J. 59: 113– 127.

33. Emanuel, P. A.,, C. Chue,, L. Kerr, and, D. Cullin. 2003. Validating the performance of biological detection equipment: the role of the federal government. Biosecur. Bio-terror. 1: 131– 137.

34. Emanuel, P. A.,, I. R. Fruchey,, A. M. Bailey,, J. L. Dang,, K. Niyogi,, J. W. Roos,, D. Cullin, and, D. C. Emanuel. 2005. Automated screening for biological weapons in homeland defense. Biosecur. Bioterror. 3: 39– 50.

35. Fabian, M. P.,, S. L. Miller,, T. Reponen, and, M. T. Hernandez. 2005. Ambient bioaerosol indices for indoor air quality assessments of flood reclamation. J. Aerosol Sci. 36: 763– 783.

36. Ferre, F. 1992. Quantitative or semi-quantitative PCR: reality versus myth. PCR Methods Applic. 2: 1– 9.

37. Flannigan, B. 1987. Mycotoxins in the air. Int. Biodeterior. 23: 73– 78.

38. Fogelmark B.,, H. Goto,, K. Yuasa,, B. Marchat, and, R. Rylander. 1992. Acute pulmonary toxicity of inhaled β-1,3-glucan and endotoxin. Agents Actions 35: 50– 56.

39. Goto, H.,, K. Yuasa, and, R. Rylander. 1994. (1-3)-β- d-Glucan in indoor air, its measurement and in vitro activity. Am. J. Ind. Med. 25: 81– 83.

40. Haugland, R. A.,, S. J. Vesper, and, L. J. Wymer. 1999. Quantitative measurement of Stachybotrys chartarum conidia using real time detection of PCR products with the TaqMan TM fluorogenic probe system. Mol. Cell. Probes 13: 329– 340.

41. Heid, C. A.,, J. Stevens,, K. J. Livak, and, P. M. Williams. 1996. Real time quantitative PCR. Genome Res. 6: 986– 994.

42. Hein, I.,, A. Lehner,, P. Rieck,, K. Klein,, E. Brandl, and, M. Wagner. 2001. Comparison of different approaches to quantify Staphylococcus aureus cells by real-time quantitative PCR and application of this technique for examination of cheese. Appl. Environ. Microbiol. 67: 3122– 3126.

43. Higgins, J. A.,, M. Cooper,, L. Schroeder-Tucker,, S. Black,, D. Miller,, J. S. Karns,, E. Manthey,, R. Breeze, and, M. L. Perdue. 2003. A field investigation of Bacillus anthracis contamination of U.S. Department of Agriculture and other Washington, D.C., buildings during the anthrax attack of October 2001. Appl. Environ. Microbiol. 69: 593– 599.

44. Hindson, B. J.,, S. B. Brown,, G. D. Marshall,, M. T. McBride,, A. J. Makarewicz,, D. M. Gutierrez,, D. K. Wolcott,, T. R. Metz,, R. S. Madabhushi,, J. M. Dzenitis, and, B. W. Colston, Jr. 2004. Development of an automated sample preparation module for environmental monitoring of biowarfare agents. Anal. Chem. 76: 3492– 3497.

45. Hirsch, S. R.,, and J. A. Sosman. 1976. A one-year survey of mold growth inside twelve homes. Ann. Allergy 36: 30– 38.

46. Hobbie, J. E.,, R. J. Daley, and, S. Jasper. 1977. Use of nucleopore filters for counting bacteria by fluorescence microscopy. Appl. Environ. Microbiol. 33: 1225– 1228.

47. Hocking, A. D.,, and J. I. Pitt. 1980. Dichloran-glycerol medium for enumeration of xerophilic fungi from low-moisture foods. Appl. Environ. Microbiol. 39: 488– 492.

48. Hunter, C. A.,, C. Grant,, B. Flannigan, and, A. F. Bravery. 1988. Mould in buildings: the air spora of domestic buildings. Int. Biodeterior. 24: 81– 101.

49. Ivnitski, D.,, D. J. O’Neil,, A. Gattuso,, R. Schlicht,, M. Calidonna, and, R. Fisher. 2003. Nucleic acid approaches for detection and identification of biological warfare and infectious agents. BioTechniques 35: 862– 869.

50. Jacobs, R. R. 1989. Airborne endotoxins: an association with occupational lung disease. Appl. Ind. Hyg. 4: 50– 56.

51. Jarvis, B. B.,, W. G. Sorenson,, E. Hintikka,, M. Nikulin,, Y. Zhou,, J. Jiang,, S. Wang,, S. Hinkley,, R. A. Etzel, and, D. Dearborn. 1998. Study of toxin production by isolates of Stachybotrys chartarum and Memnoniella echinata isolated during a study of pulmonary hemosiderosis in infants. Appl. Environ. Microbiol. 64: 3620– 3625.

52. Jensen, P. A.,, B. Lighthart,, A. J. Mohr, and, B. T. Shaffer. 1994. Instrumentation used with microbial bioaerosol, p. 226–284. In B. Lighthart and, A. J. Mohr (ed.), Atmospheric Microbial Aerosols: Theory and Applications. Chapman and Hall, New York, N.Y.

53. Johanning, E.,, R. Biagini,, D. Hull,, P. Morey,, B. Jarvis, and, P. Landsbergis. 1996. Health and immunology study following exposure to toxigenic fungi ( Stachybotrys chartarum) in a water-damaged office environment. Int. Arch. Occup. Environ. Health 68: 207– 218.

54. Josephson, K. L.,, C. P. Gerba, and, I. L. Pepper. 1993. Polymerase chain reaction detection of nonviable bacterial pathogens. Appl. Environ. Microbiol. 59: 3513– 3515.

55. Korpi, A.,, A.-L. Pasanen, and, P. Pasanen. 1998. Volatile compounds originating from mixed microbial cultures on building materials under various humidity conditions. Appl. Environ. Microbiol. 64: 2914– 2919.

56. Lange, J. L.,, P. S. Thorne, and, N. Lynch. 1997. Application of flow cytometry and fluorescent in situ hybridization for assessment of exposures to airborne bacteria. Appl. Environ. Microbiol. 63: 1557– 1563.

57. Leenders, A. C. A. P.,, A. Van Belkum,, M. Behrendt,, A. Luijendijk, and, H. A. Verbrugh. 1999. Density and molecular epidemiology of Aspergillus in air and relationship to outbreaks of Aspergillus infection. J. Clin. Microbiol. 37: 1752– 1757.

58. Leser, T. D.,, M. Boye, and, N. B. Hendriksen. 1995. Survival and activity of Pseudomonas sp. strain B13(FR1) in a marine microcosm determined by quantitative PCR and an rRNA-targeting probe and its effect on the indigenous bacterioplankton. Appl. Environ. Microbiol. 61: 1201– 1207.

59. Levetin, E. 1995. Fungi, p. 87–120. In H. A. Burge (ed.), Bioaerosols. Lewis Publishers, Boca Raton, Fla.

60. Loeffler, J.,, N. Henke,, H. Hebart,, D. Schmidt,, L. Hagmeyer,, U. Schumacher, and, H. Einsele. 2000. Quantification of fungal DNA by using fluorescence resonance energy transfer and the Light Cycler system. J. Clin. Microbiol. 38: 586– 590.

61. Loy, A.,, M. Horn, and, M. Wagner. 2003. probeBase: an online resource for rRNA-targeted oligonucleotide probes. Nucleic Acids Res. 31: 514– 516.

62. Luczynska, C. M.,, Y. Li,, M. D. Chapman, and, T. A. E. Platts-Mills. 1990. Airborne concentration and particle size distribution of allergen derived from domestic cats (Felis domesticus): measurements using cascade impactor, liquid impinger and a two site monoclonal antibody assay for Fel d I. Am. Rev. Respir. Dis. 141: 361– 367.

63. Luo, G.,, and T. G. Mitchell. 2002. Rapid identification of pathogenic fungi directly from cultures by using multiplex PCR. J. Clin. Microbiol. 40: 2860– 2865.

64. Macher, J. M. 1989. Positive-hole correction of multiple-jet impactors for collecting viable microorganisms. Am. Ind. Hyg. Assoc. J. 50: 561– 568.

65. Madelin, T. M.,, and M. F. Madelin. 1995. Biological analysis of fungi and associated molds, p. 361–386. In C. S. Cox and, C. M. Wathes (ed.), Bioaerosols Handbook. CRC Press, Boca Raton, Fla.

66. McBride, M. T.,, D. Masquelier,, B. J. Hindson,, A. J. Makarewicz,, S. Brown,, K. Burris,, T. Metz,, R. G. Langlois,, K. W. Tsang,, R. Bryan,, D. A. Anderson,, K. S. Venkateswaran,, F. P. Milanovich, and, B. W. Colston, Jr. 2003. Autonomous detection of aerosolized Bacillus anthracis and Yersinia pestis. Anal. Chem. 75: 5293– 5299.

67. Miller, J. D.,, A. M. Laflamme,, Y. Sobol,, P. Lafontaine, and, R. Greenhalgh. 1988. Fungi and fungal products in some Canadian houses. Int. Biodeterior. 24: 103– 120.

68. Miller, J. D.,, and J. C. Young. 1997. The use of ergosterol to measure exposure to fungal propagules in indoor air. Am. Ind. Hyg. Assoc. J. 58: 39– 43.

69. Milton, D. K. 1995. Endotoxin, p. 77–86. In H. A. Burge (ed.), Bioaerosols. CRC Press, Boca Raton, Fla.

70. Milton, D. K.,, R. J. Gere,, H. A. Feldman, and, I. A. Greaves. 1990. Endotoxin measurement: aerosol sampling and application of a new Limulus method. Am. Ind. Hyg. Assoc. J. 51: 331– 337.

71. Morring, K. L.,, W. G. Sorenson, and, M. D. Attfield. 1983. Sampling for airborne fungi: a statistical comparison of media. Am. Ind. Hyg. Assoc. J. 44: 662– 664.

72. Myatt, T. A.,, S. L. Johnston,, Z. Zuo,, M. Wand,, T. Kebadze,, S. Rudnick, and, D. K. Milton. 2004. Detection of airborne rhinovirus and its relation to outdoor air supply in office environments. Am. J. Respir. Crit. Care Med. 169: 1187– 1190.

73. Nogva, H. K.,, and D. Lillehaug. 1999. Detection and quantification of Salmonella in pure cultures using 5′-nuclease polymerase chain reaction. Int. J. Food Microbiol. 51: 191– 196.

74. Olsson, M.,, C. Lidman,, S. Latouche,, A. Björkman,, P. Roux,, E. Linder, and, M. Wahlgren. 1998. Identification of Pneumocystis carinii f. sp. hominis gene sequences in filtered air in hospital environments. J. Clin. Microbiol. 36: 1737– 1740.

75. Orlando, C.,, P. Pinzani, and, M. Pazzagli. 1998. Developments in quantitative PCR. Clin. Chem. Lab. Med. 36: 255– 269.

76. Palmgren, U.,, G. Strom,, G. Blomquist, and, P. Malmberg. 1986. Collection of airborne micro-organisms on Nuclepore filters, estimation and analysis—CAMNEA method. J. Appl. Bacteriol. 61: 401– 406.

77. Panaccione, D. G.,, and C. M. Coyle. 2005. Abundant respirable ergot alkaloids from the common airborne fungus Aspergillus fumigatus. Appl. Environ. Microbiol. 71: 3106– 3111.

78. Pasanen, A.,, M. Nikulin,, M. Tuomainen,, S. Berg,, P. Parikka, and, E. Hintikka. 1993. Laboratory experiments on membrane filter sampling of airborne mycotoxins produced by Stachybotrys atra corda. Atmos. Environ. 27A: 9– 13.

79. Picard, C.,, C. Ponsonnet,, E. Paget,, X. Nesme, and, P. Simonet. 1992. Detection and enumeration of bacteria in soil by direct DNA extraction and polymerase chain reaction. Appl. Environ. Microbiol. 58: 2717– 2722.

80. Platts-Mills, T. A. E.,, W. R. Thomas,, R. C. Aalbersee,, D. Vervloet, and, M. D. Chapman. 1992. Dust mite allergens and asthma: report of a second international workshop. J. Allergy Clin. Immunol. 89: 1046.

81. Prigione, V.,, G. Lingua, and, V. Filipello Marchisio. 2004. Development and use of flow cytometry for detection of airborne fungi. Appl. Environ. Microbiol. 70: 1360– 1365.

82. Reesley, M.,, M. Miller, and, K. F. Nielsen. 2003. Quantifying mold biomass on gypsum board: comparison of ergosterol and beta- N-acetylhexosaminidase as mold biomass parameters. Appl. Environ. Microbiol. 69: 3996– 3998.

83. Rylander, R. 1999. Indoor air-related effects and airborne (1→3)-β- d-glucan. Environ. Health Perspect. 107: 501– 503.

84. Saiki, R. K.,, S. Scharf,, F. Faloona,, K. B. Mullis,, G. T. Horn,, H. A. Erlich, and, N. Arnheim. 1985. Enzymatic amplification of β-globin genomic sequence and restriction site analysis for diagnosis of sickle cell anemia. Science 230: 1350– 1354.

85. Sawyer, M. H.,, C. J. Chamberlain,, Y. N. Wu,, N. Aintablian, and, M. R Wallace. 1994. Detection of varicella-zoster virus DNA in air samples from hospital rooms. J. Infect. Dis. 169: 91– 94.

86. Sebastian, A.,, and L. Larsson. 2003. Characterization of the microbial community in indoor environments: a chemical-analytical approach. Appl. Environ. Microbiol. 69: 3103– 3109.

87. Smid, T.,, E. Schokkin,, J. S. M. Boleij, and, D. Heederik. 1989. Enumeration of viable fungi in occupational environments: a comparison of samplers and media. Am. Ind. Hyg. Assoc. J. 50: 235– 239.

88. Solomon, W. R. 1975. Assessing fungus prevalence in domestic interiors. J. Allergy Clin. Immunol. 56: 235– 242.

89. Sorenson, W. G.,, D. G. Frazer,, B. B. Jarvis,, J. Simpson, and, V. A. Robinson. 1987. Trichothecene mycotoxins in aerosolized conidia of Stachybotrys atra. Appl. Environ. Microbiol. 53: 1370– 1375.

90. Stärk, K. D. C.,, J. Nicolet, and, J. Frey. 1998. Detection of Mycoplasma hypopneumoniae by air sampling with a nested PCR assay. Appl. Environ. Microbiol. 64: 543– 548.

91. Strachan, D. P.,, B. Flannigan,, E. M. McCabe, and, F. McGarry. 1990. Quantification of airborne moulds in the homes of children with and without wheeze. Thorax 45: 382– 387.

92. Tuomi, T.,, K. Reijula,, T. Johnsson,, K. Hemminki,, E.-L. Hintikka,, O. Lindroos,, S. Kalso,, P. Koukila-Kähkölä,, H. Mussalo-Rauhamaa, and, T. Haahtela. 2000. Mycotoxins in crude building materials from water-damaged buildings. Appl. Environ. Microbiol. 66: 1899– 1904.

93. Vadrot, C.,, V. Bex,, A. Mouilleseaux,, F. Squinazi, and, J.-C. Darbord. 2004. Detection of Mycobacterium tuberculosis complex by PCR in hospital air samples. J. Hosp. Infect. 58: 262– 267.

94. Verhoeff, A. P.,, J. H. van Wijnen,, J. S. M. Boleij,, B. Brunekreef,, E. S. van Reenen-Hoekstra, and, R. A. Samson. 1990. Enumeration and identification of airborne viable mould propagules in houses. Allergy 45: 275– 284.

95. Vesper, S.,, D. G. Dearborn,, I. Yike,, T. Allan,, J. Sobolewski,, S. F. Hinkley,, B. B. Jarvis, and, R. A. Haugland. 2000. Evaluation of Stachybotrys chartarum in the house of an infant with pulmonary hemorrhage: quantitative assessment before, during, and after remediation. J. Urban Health 77: 68– 84.

96. Walter, M. V.,, B. Marthi,, V. P. Fieland, and, L. M. Ganio. 1990. Effect of aerosolization on subsequent bacterial survival. Appl. Environ. Microbiol. 56: 3468– 3472.

97. Walters, M.,, D. Milton,, L. Larsson, and, T. Ford. 1994. Airborne environmental endotoxin: a cross validation of sampling and analysis techniques. Appl. Environ. Micro-biol. 60: 996– 1005.

98. Weimer, B. C.,, M. K. Walsh,, C. Beer,, R. Koka, and, X. Wang. 2001. Solid-phase capture of proteins, spores, and bacteria. Appl. Environ. Microbiol. 67: 1300– 1307.

99. White, T. J.,, T. Bruns,, S. Lee, and, J. Taylor. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics, p. 315–322. In M. A. Innis,, D. H. Gelfand,, J. J. Sninsky, and, T. J. White (ed.), PCR Protocols: a Guide to Methods and Applications. Academic Press, San Diego, Calif.

100. Wilson, I. G. 1997. Inhibition and facilitation of nucleic acid amplification. Appl. Environ. Microbiol. 63: 3741– 3751.

101. Wu, Z.,, X. R. Wang, and, G. Blomquist. 2002. Evaluation of PCR primers and PCR conditions for specific detection of common airborne fungi. J. Environ. Monit. 4: 377– 382.

102. Yike, I.,, T. Allan,, W. G. Sorenson, and, D. G. Dearborn. 1999. Highly sensitive protein translation assay for trichothecene toxicity in airborne particulates: comparison with cytotoxicity assays. Appl. Environ. Microbiol. 65: 88– 94.

103. Zeng, Q.-Y.,, S.-O. Westermark,, Å. Rasmuson-Lestander, and, X.-R. Wang. 2004. Detection and quantification of Wallemia sebi in aerosols by real-time PCR, conventional PCR, and cultivation. Appl. Environ. Microbiol. 70: 7295– 7302.