1. Utility of PCR amplification and DNA microarray hybridization of 16S rDNA for rapid diagnosis of bacteremia associated with hematological diseases
- Author
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Eiju Negoro, Mitsunobu Shimadzu, Akira Yoshida, Shinji Kishi, Yoshimasa Urasaki, Katsunori Tai, Kazutaka Takagi, Satoshi Ikegaya, Takanori Ueda, Hiromichi Iwasaki, and Takahiro Yamauchi
- Subjects
DNA, Bacterial ,Microbiology (medical) ,Microarray ,Rapid diagnosis ,16S ribosomal DNA ,Bacteremia ,Biology ,DNA, Ribosomal ,Polymerase Chain Reaction ,Sensitivity and Specificity ,law.invention ,Nucleic acid thermodynamics ,law ,RNA, Ribosomal, 16S ,Gene duplication ,medicine ,Hematological disease ,Humans ,Blood culture ,Ribosomal DNA ,Polymerase chain reaction ,DNA Primers ,Oligonucleotide Array Sequence Analysis ,medicine.diagnostic_test ,Bacteria ,DNA microarray ,Gene Amplification ,Nucleic Acid Hybridization ,General Medicine ,medicine.disease ,Virology ,Molecular biology ,Hematologic Diseases ,Infectious Diseases - Abstract
Summary Objectives The rapid diagnosis of bacteremia is crucial for patient management including the choice of antimicrobial therapy, especially in cases of hematological disease, because neutropenia occurs frequently during antineoplastic chemotherapy or disease progression. We describe a rapid detection and identification system that uses universal PCR primers to amplify a variable region of bacterial 16S ribosomal DNA (rDNA), followed by DNA microarray hybridization. Methods Probes for 72 microorganisms including most causal clinical pathogens were spotted onto a microarray plate. The DNA microarray and conventional methods of identification were applied to 335 cultures from patients with hematological diseases. Results Forty-one samples (12.2%) tested positive by conventional blood culture test in a few days, while 40 cases (11.9%) were identified by the new method within 24h. The sensitivity and specificity of this new method were 93% and 99%, respectively, compared with conventional blood culture testing. Conclusions PCR combined with a DNA microarray is useful for the management of febrile patients with hematological diseases.
- Published
- 2013
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