DNA probe technology for rapid detection of Haemophilus influenzae in clinical specimens

Malouin, François; Bryan, L. E.; Shewciw, P; Douglas, James T.; Li, D; Elzen, H van den; Lapointe, Jean-Rock

doi:10.1128/jcm.26.10.2132-2138.1988

Cited by 16 publications

(3 citation statements)

References 17 publications

(29 reference statements)

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“…Numerous studies have shown the accutracy of DNA probes for the direct identification of a variety of bacteria in clinical specimens (8,9,13,15,18,20) and as culture confirmation tests (5,12,14,17,22,26,27). This report assesses the potential usefulness of commercially available DNA probe kits for the direct, rapid identification of bacteria from positive blood culture broths.…”

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confidence: 98%

Direct identification of bacterial isolates in blood cultures by using a DNA probe

Davis

Fuller

1991

J Clin Microbiol

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This study involved the rapid, direct identification of Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, Enterococcus sp., and Streptococcus agalactiae from positive blood culture bottles (BACTEC, Johnston Laboratories, Inc.) by using the AccuProbe (Gen-Probe, San Diego, Calif.) culture confirmation test. This method uses a chemiluminescent DNA probe that detects the rRNA of the target organisms. The manufacturer's instructions were modified to use a pellet of bacteria made directly from positive blood culture broth rather than a colony from an agar plate. Two separate procedures of selective centrifugation were employed in order to obtain the pellet. The first utilized a routine clinical centrifuge and a large volume of broth (10 to 12 ml) from the blood culture bottle. The second method used a microcentrifuge and less volume (1 to 1.5 ml).-A total of 196 clinical specimens taken directly from positive blood culture broths were correctly identified by AccuProbe from pellets made by using the clinical centrifuge technique, while 166 clinical specimens used as negative controls failed to show hybridization. The microcentrifuge technique for obtaining pellets was performed on 105 patient specimens, and all were correctly identified. When combined with the microcentrifuge technique for pellet preparation, the AccuProbe test has several advantages: (i) direct identification of bacteria from blood culture broths, (ii) rapid turnaround time (30 min), (iii) simplicity of the procedure, and (iv) relative low cost.

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confidence: 98%

Direct identification of bacterial isolates in blood cultures by using a DNA probe

Davis

Fuller

1991

J Clin Microbiol

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“…In a previous study it has been shown that H. influenzae displays a characteristic temperature-sensitive PBP3 with a reduced penicillin-binding activity at 42°C ( Malouin et al, 1988 ). Taking into account the fact that in E. coli , the inhibition of PBP1b alone or combined with that of PBP1a by β-lactams causes the rapid lysis of bacteria cells, it is more than likely that an additional inhibitory effect linked to the temperature-sensitive PBP3 should be considered for the increased imipenem sensitivity of GE47 and GE88 strains grown at 42°C.…”

Section: Discussionmentioning

confidence: 99%

Transcriptional Modulation of Penicillin-Binding Protein 1b, Outer Membrane Protein P2 and Efflux Pump (AcrAB-TolC) during Heat Stress Is Correlated to Enhanced Bactericidal Action of Imipenem on Non-typeable Haemophilus influenzae

et al. 2018

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Objective: The purpose of the present study was to investigate the penicillin binding proteins (PBPs), drug influx and efflux modulations during heat stress and their effects on the bactericidal action of imipenem on non-typeable Haemophilus influenzae (NTHi).Methods: The two NTHi clinical isolates (GE47 and GE88, imipenem MICs by E-test > 32 μg/mL) examined in this study were collected at Geneva University Hospitals. The imipenem killing activity was assessed after incubation of the NTHi strains at either 37 or 42°C for 3 h with increasing concentrations of imipenem. The detection of PBPs was carried out by Bocillin-FL. Global transcriptional changes were monitored by RNA-seq after pre-incubation of bacterial cells at either 37 or 42°C, and the expression levels of relevant target genes were confirmed by qRT-PCR.Results: Quantitation of NTHi viable cells after incubation with 0.25 μg/mL of imipenem for 3 h revealed more than a twofold decrease in GE47 and GE88 viable cells at 42°C as compared to 37°C. Transcriptome analysis showed that under heat stress conditions, there were 141 differentially expressed genes with a | log2(fold change)| > 1, including 67 up-regulated and 74 down-regulated genes. The expression levels of ponB (encoding PBP1b) and acrR (regulator of AcrAB-TolC efflux pump) were significantly increased at 42°C. In contrast, the transcript levels of ompP2 (encoding the outer membrane protein P2) and acrB gene (encoding AcrB) were significantly lower under heat stress condition.Conclusion: This study shows that the transcriptional modulation of ponB, ompP2, acrR, and acrB in the heat stress response is correlated to enhanced antimicrobial effects of imipenem on non-typeable H. influenzae.

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“…Up to now, the main emphasis in constructing such DNA probes was to have quick and reliable tools to identify pathogenic material in clinical diagnosis. Thus, DNA probes have already been used for the identification of Plasmodium falciparum (1), Yersinia enterocolitica (6), Salmonella typhi (14), Bacillus subtilis (7), Haemophilus influenzae (10), and other microorganisms and of DNA viruses (2,16,17) and RNA viruses (4,9). For the genus Lactobacillus, a probe for L. curvatus (12), which is specifically associated with spoilage of vacuum-packed meats (13), has been reported.…”

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confidence: 99%

DNA Probe for Lactobacillus delbrueckii

Delley

Mollet

Hottinger

1990

Appl Environ Microbiol

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From a genomic DNA library of Lactobacilus delbrueckii subsp. bulgaricus, a clone was isolated which complements a leucine auxotrophy of an Escherichia coli strain (GE891). Subsequent analysis of the clone indicated that it could serve as a specific DNA probe. Dot-blot hybridizations with over 40 different Lactobacillus strains showed that this clone specifically recognizes L. delbrueckii subsp. delbrueckii, bulgaricus, and lactis. The sensitivity of the method was tested by using an a-32P-labeled DNA probe.

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DNA probe technology for rapid detection of Haemophilus influenzae in clinical specimens

Cited by 16 publications

References 17 publications

Direct identification of bacterial isolates in blood cultures by using a DNA probe

Direct identification of bacterial isolates in blood cultures by using a DNA probe

Transcriptional Modulation of Penicillin-Binding Protein 1b, Outer Membrane Protein P2 and Efflux Pump (AcrAB-TolC) during Heat Stress Is Correlated to Enhanced Bactericidal Action of Imipenem on Non-typeable Haemophilus influenzae

DNA Probe for Lactobacillus delbrueckii

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