Multiplexed Identification of Blood-Borne Bacterial Pathogens by Use of a Novel 16S rRNA Gene PCR-Ligase Detection Reaction-Capillary Electrophoresis Assay

Pingle, Maneesh; Granger, Kathleen; Feinberg, Philip B.; Shatsky, Rebecca; Sterling, Bram; Rundell, Mark S.; Spitzer, Eric D.; Larone, Davise H.; Golightly, Linnie M.; Barany, Francis

doi:10.1128/jcm.00226-07

Cited by 43 publications

(49 citation statements)

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“…The hybridization time may be reduced even further to 20 min using the slide chamber method, leading to a result in only 60 min. This is faster than biochemical methods and other molecular identification methods [1,[7][8][9], including the previously described Acinetobacter-FISH procedures [18,19]. The newly developed probe may be used on multi-well glass slides in combination with a broader panel of previously published probes including probes for non-fermentative bacteria and Enterobacteriaceae [10,11].…”

Section: Discussionmentioning

confidence: 99%

“…These systems are quite reliable but time-consuming [6]. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and molecular methods such as NAAT (nucleic acid amplification technique) [1], amplified ribosomal DNA restriction analysis [7], 16S rRNA gene PCR-ligase detection reaction-capillary electrophoresis assays [8] and DNA microarrays [9] allow for rapid and reliable identification but are expensive, require sophisticated equipment and are not generally available in routine clinical laboratories. Fluorescence in situ hybridization (FISH) is an alternative molecular method for rapid identification of bacteria that is more cost-effective than automated NAAT platforms and requires less technical expertise to perform than most other available molecular methods.…”

Section: Introductionmentioning

confidence: 99%

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Rapid identification ofAcinetobacterspp. by fluorescencein situhybridization (FISH) from colony and blood culture material

Frickmann¹,

Essig²,

Hagen³

et al. 2011

European Journal of Microbiology and Immunology

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Multi-drug-resistant strains of the Acinetobacter baumannii complex cause nosocomial infections. Rapid identification of Acinetobacter spp. is desirable in order to facilitate therapeutic or hygiene decisions. We evaluated a newly designed DNA probe that can be used under standard conditions in both a microwave oven and a slide chamber for the rapid identification of Acinetobacter spp. by fluorescence in situ hybridization (FISH). Using FISH, the new probe correctly identified 81/81 Acinetobacter spp. isolates and excluded 109/109 tested non-target organisms from agar culture. Furthermore, the new probe correctly identified 7/7 Acinetobacter spp. in 214 blood cultures determined to contain Gram-negative bacteria by Gram staining. Using either the microwave oven or slide chamber technique, the new probe was able to identify Acinetobacter spp. in 100% of the samples tested. FISH used in conjunction with our newly designed probe provides an easy, cheap, precise, and rapid method for the preliminary identification of Acinetobacter spp., especially in laboratories where more sophisticated methods like matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) are not available.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Rapid identification ofAcinetobacterspp. by fluorescencein situhybridization (FISH) from colony and blood culture material

Frickmann¹,

Essig²,

Hagen³

et al. 2011

European Journal of Microbiology and Immunology

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“…and detect as few as 100 A. baumannii cells per mL blood (Lin et al, 2008). A highthroughput method based on PCR-ligase reaction and capillary electrophoresis was described which could detect A. baumannii in blood (Pingle et al, 2007). These methods are faster than PCR-DGGE but may be limited to detecting only known or previously described sequences.…”

Section: Detection Methods That Do Not Require Cultivationmentioning

confidence: 99%

Safety Assessment of Transgenic Organisms, Volume 4

2010

Harmonisation of Regulatory Oversight in Biotechnology

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io s a fe t y nv ir on me nt ag ri cu lt ur e bi os af et y g r ic u lt u r e e n vi r o n m e n t gr ic ul tu re bi os af et y en vi ro nm en t n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o io sa fe ty en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en v io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t nv ir on me nt ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm e g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a f gr ic ul tu re bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t a n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u io sa fe ty en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m nv ir on me nt ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a f gr ic ul tu re bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t a n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u io sa fe ty en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e nv ir on me nt ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y e g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a fe t y a g r ic u lt u r e e n vi r o n m e n t b io s a f ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et y en vi ro nm en t ag ri cu lt ur e bi os af et...

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“…The LDR products are separated in capillaries and the ligation occurrence is visible due to the band-shift. The use of various probe sets simultaneously is still possible and the ligated probes can be separated with a one base pair resolution [17,46]. This approach can be cost effective, but when the simultaneous visualization of many different DNA species or a higher sensitivity is needed UAs are more convenient.…”

Section: Ligation Detection Reaction-universal Arrays (Ldr-ua)mentioning

confidence: 99%

Potentials and limitations of molecular diagnostic methods in food safety

Lauri

Mariani

2008

Genes Nutr

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Molecular methods allow the detection of pathogen nucleic acids (DNA and RNA) and, therefore, the detection of contamination in food is carried out with high selectivity and rapidity. In the last 2 decades molecular methods have accompanied traditional diagnostic methods in routine pathogen detection, and might replace them in the upcoming future. In this review the implementation in diagnostics of four of the most used molecular techniques (PCR, NASBA, microarray, LDR) are described and compared, highlighting advantages and limitations of each of them. Drawbacks of molecular methods with regard to traditional ones and the difficulties encountered in pathogen detection from food or clinical specimen are also discussed. Moreover, criteria for the choice of the target sequence for a secure detection and classification of pathogens and possible developments in molecular diagnostics are also proposed.

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Multiplexed Identification of Blood-Borne Bacterial Pathogens by Use of a Novel 16S rRNA Gene PCR-Ligase Detection Reaction-Capillary Electrophoresis Assay

Abstract: We have developed a novel high-throughput PCR-ligase detection reaction-capillary electrophoresis (PCR-LDR

Cited by 43 publications

References 65 publications

Rapid identification ofAcinetobacterspp. by fluorescencein situhybridization (FISH) from colony and blood culture material

Rapid identification ofAcinetobacterspp. by fluorescencein situhybridization (FISH) from colony and blood culture material

Safety Assessment of Transgenic Organisms, Volume 4

Potentials and limitations of molecular diagnostic methods in food safety

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