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Chapter 2 : Conventional Blood Culture Methods

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Conventional Blood Culture Methods, Page 1 of 2

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Abstract:

Chapter 1 took us on a time-travel journey spanning the evolution of blood cultures. All the experimentation and discovery since the 17th century have led to a common understanding of which microorganisms cause bloodstream infections and what essential nutrition is required to grow and recover them from blood. Even today, blood culture remains the best option to determine microbial causes of sepsis. Detection of microbial nucleic acids in blood by PCR or sequencing may not represent the presence of viable organisms (see chapter 13 on the blood microbiome). Often, manual culture methods are used as a reference standard for newer iterative protocols like current-generation automated blood culture systems ( ). While the routine use of manual blood culture methods in modern diagnostic microbiology laboratories in 2017 is no longer commonplace, the practice remains quite prevalent in the developing world where purchase and maintenance of automated blood culture instruments are both unaffordable and impractical. But in the developed world, manual blood culture protocols often serve as a backup plan if automated systems experience problems. The comedian Mitch Hedberg once said, “An escalator can never break: it can only become stairs.” The same holds true for automated blood culture systems. Regardless of how blood cultures are performed, however, the same set of parameters must be attained to ensure optimal microbial recovery. The list of best practice benchmarks includes the following and can also be found in chapter 10:

Citation: Atkinson-Dunn R, Dunne W. 2017. Conventional Blood Culture Methods, p 21-38. In Dunne, Jr. W, Burnham C (ed), The Dark Art of Blood Cultures. ASM Press, Washington, DC. doi: 10.1128/9781555819811.ch2
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Figures

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Figure 1

Septi-Chek blood culture system by BBL. This is a two-piece manual culture system designed for culture of blood. The system consists of a broth bottle available in five formulations and two media fill volumes with or without antimicrobial removal resins and an upper slide unit consisting of paddles with solid agar media. Blood is introduced into the broth, and the slide unit is attached to the top. Periodically, the blood/broth mixture is inverted such that it washes over the surface of the agar paddles. It is then incubated in the upright position and the bottles are inspected for growth in the broth and colonies on the agar surface. Image used with permission from BD Biosciences.

Citation: Atkinson-Dunn R, Dunne W. 2017. Conventional Blood Culture Methods, p 21-38. In Dunne, Jr. W, Burnham C (ed), The Dark Art of Blood Cultures. ASM Press, Washington, DC. doi: 10.1128/9781555819811.ch2
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Image of Figure 2
Figure 2

Oxoid Signal blood culture system by Oxoid. Also a two-piece manual blood culture system consisting of a broth-containing bottle and a growth indicator device that is attached to the top of the bottle. Organism growth generates positive pressure in the bottle that displaces a volume of the blood-broth mixture into the detection chamber. Image used with permission from BD Biosciences.

Citation: Atkinson-Dunn R, Dunne W. 2017. Conventional Blood Culture Methods, p 21-38. In Dunne, Jr. W, Burnham C (ed), The Dark Art of Blood Cultures. ASM Press, Washington, DC. doi: 10.1128/9781555819811.ch2
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Figure 3

Blood culture bottle metal cap crimping device.

Citation: Atkinson-Dunn R, Dunne W. 2017. Conventional Blood Culture Methods, p 21-38. In Dunne, Jr. W, Burnham C (ed), The Dark Art of Blood Cultures. ASM Press, Washington, DC. doi: 10.1128/9781555819811.ch2
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References

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1. Baron EJ,, Weinstein MP,, Dunne WM Jr,, Yagupsky P,, Welch DF,, Wilson DM,. 2005. Cumitech 1C, Blood Cultures IV. Baron EJ, Coordinating ed. ASM Press, Washington, DC.
2. Wilson ML,, Mitchell M,, Morris AJ,, Murray PR,, Reimer LG,, Reller LB,, Towns M,, Weinstein MP,, Wellstood SA,, Dunne WM Jr,, Jerris RC,, Welch DF. 2007. Principles and Procedures for Blood Cultures; Approved Guideline M47-A. CLSI, Wayne, PA.
3. Riedel S,, Bourbeau P,, Swartz B,, Brecher S,, Carroll KC,, Stamper PD,, Dunne WM,, McCardle T,, Walk N,, Fiebelkorn K,, Sewell D,, Richter SS,, Beekmann S,, Doern GV. 2008. Timing of specimen collection for blood cultures from febrile patients with bacteremia. J Clin Microbiol 46:13811385.
4. Dunne WM Jr,, LaRocco M,. 2001. Blood culture systems, p 189209. In Cimolai N (ed), Laboratory Diagnosis of Bacterial Infections. Marcel-Dekker, New York, NY.
5. Schaffner J,, Chochua S,, Kourbatova EV,, Barragan M,, Wang YF,, Blumberg HM,, del Rio C,, Walker HK,, Leonard MK. 2009. High mortality among patients with positive blood cultures at a children’s hospital in Tbilisi, Georgia. J Infect Dev Ctries 3:267272.
6. Campbell J,, Washington JA II. 1980. Evaluation of the necessity for routine terminal subcultures of previously negative blood cultures. J Clin Microbiol 12:576578.
7. Gill VJ. 1981. Lack of clinical relevance in routine terminal subculturing of blood cultures. J Clin Microbiol 14:116118.
8. Blazevic DJ,, Stemper JE,, Matsen JM. 1974. Comparison of macroscopic examination, routine gram stains, and routine subcultures in the initial detection of positive blood cultures. Appl Microbiol 27:537539.
9. Doern GV,, Brueggemann AB,, Dunne WM,, Jenkins SG,, Halstead DC,, McLaughlin JC. 1997. Four-day incubation period for blood culture bottles processed with the Difco ESP blood culture system. J Clin Microbiol 35:12901292.
10. Han XY,, Truant AL. 1999. The detection of positive blood cultures by the AccuMed ESP-384 system: the clinical significance of three-day testing. Diagn Microbiol Infect Dis 33:16.
11. Castaneda MR. 1947. A practical method for routine blood cultures in brucellosis. Proc Soc Exp Biol Med 64:114115.
12. Murray PR,, Niles AC,, Heeren RL,, Curren MM,, James LE,, Hoppe-Bauer JE. 1988. Comparative evaluation of the oxoid signal and Roche Septi-Chek blood culture systems. J Clin Microbiol 26:25262530.
13. Daley C,, Lim I,, Modra J,, Wilkinson I. 1990. Comparative evaluation of nonradiometric BACTEC and improved oxoid signal blood culture systems in a clinical laboratory. J Clin Microbiol 28:15861590.
14. Rohner P,, Pepey B,, Auckenthaler R. 1995. Comparison of BacT/Alert with Signal blood culture system. J Clin Microbiol 33:313317.
15. Weinstein MP,, Mirrett S,, Wilson ML,, Harrell LJ,, Stratton CW,, Reller LB. 1991. Controlled evaluation of BACTEC Plus 26 and Roche Septi-Chek aerobic blood culture bottles. J Clin Microbiol 29:879882.
16. Welby PL,, Zusag TM,, Storch GA. 1992. Comparison of the Bactec Peds Plus pediatric blood culture vial with Roche pediatric Septi-Chek for blood cultures from pediatric patients. J Clin Microbiol 30:13611362.
17. Culture Media Special Interest Group for the Australian Society for Microbiology. 2012. Guidelines for Assuring Quality of Medical Microbiological Culture Media. The Australian Society for Microbiology, Melbourne, Victoria, Australia.

Tables

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Table 1

Sampling plan for sterility control of blood culture bottles from Australian Guidelines ( )

Citation: Atkinson-Dunn R, Dunne W. 2017. Conventional Blood Culture Methods, p 21-38. In Dunne, Jr. W, Burnham C (ed), The Dark Art of Blood Cultures. ASM Press, Washington, DC. doi: 10.1128/9781555819811.ch2
Generic image for table
Untitled

Strains used as positive control

Citation: Atkinson-Dunn R, Dunne W. 2017. Conventional Blood Culture Methods, p 21-38. In Dunne, Jr. W, Burnham C (ed), The Dark Art of Blood Cultures. ASM Press, Washington, DC. doi: 10.1128/9781555819811.ch2

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