Direct identification of <i>Yersinia enterocolitica</i> in blood by polymerase chain reaction amplification

Feng, Peter; Keasler, S P; Hill, Walter E.

doi:10.1046/j.1537-2995.1992.32993110759.x

Cited by 67 publications

(31 citation statements)

References 19 publications

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“…Sensitive methods are particularly necessary to detect pathogenic Y. enterocolitica in asymptomatic carriers, e.g., to study possible animal reservoirs for this pathogen. Rapid and sensitive methods are also needed to detect small numbers of Y. enterocolitica organisms and other bacteria in blood units used for transfusion or in asymptomatic blood donors (40,135).…”

Section: Clinical Samplesmentioning

confidence: 99%

Low Occurrence of Pathogenic Yersinia enterocolitica in Clinical, Food, and Environmental Samples: a Methodological Problem

2003

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While Yersinia enterocolitica is an important pathogen, which can cause yersiniosis in humans and animals, its epidemiology remains obscure. The pig is the major reservoir of pathogenic Y. enterocolitica of bioserotype 4/O:3, the most common type found in humans. Y. enterocolitica is thought to be a significant food-borne pathogen, although pathogenic isolates have seldom been recovered from foods. The low isolation rate of this pathogenic bacterium in natural samples, including clinical, food, and environmental samples, may be due to the limited sensitivity of culture methods. During the last decade, numerous DNA-based methods, such as PCR and colony hybridization assays, have been designed to detect pathogenic Y. enterocolitica in natural samples more rapidly and with better sensitivity than can be achieved by culture methods. In addition, the occurrence of pathogenic Y. enterocolitica in natural samples is clearly higher with PCR than with culture methods. The methods available for detection of pathogenic Y. enterocolitica in natural samples are reviewed in this article

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Section: Clinical Samplesmentioning

confidence: 99%

Low Occurrence of Pathogenic Yersinia enterocolitica in Clinical, Food, and Environmental Samples: a Methodological Problem

2003

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“…Traditional polymerase chain reaction (PCR) assay is an effective way to detect and identify the organisms in animals and foods, including pathogenic Y. enterocolitica [4,5,6,7,8]. Mutiplex PCR was used to confirm the presumptive Y. enterocolitica isolates cultured from pigs [9].…”

Section: Introductionmentioning

confidence: 99%

Clinical Isolation and Characterization of Yersinia enterocolitica in China Using Real-Time PCR and Culture Method

et al. 2007

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Objective: To carry out a preliminary epidemiological investigation of Yersinia enterocolitica from a clinical perspective in China and to further validate the consistency of real-time polymerase chain reaction (RT-PCR) and culture method. Methods: 700 stool samples were randomly and continuously collected from in- and outpatients with diarrhea, which were detected by both the culture method and RT-PCR. Results: All 52 positive samples were successfully detected by both RT-PCR and the culture method. The total incidence of yersiniosis was about 7.4%, which was statistically higher than before (0.65%) by the Mann-Whitney U test. The morbidity was not significant between males and females (U = 0.354, p > 0.05) by the Mann-Whitney U test whereas it was significant between children and adults (U = 2.06, p < 0.05) as well as between the autumn-winter and spring-summer (U = 2.04, p < 0.05) months. Moreover, the constituent ratio of children and adults with yersiniosis in different systems was not significant by the McNemer test while it was significant between children and adults in the incidence of extraintestinal infection (U = 2.51, p < 0.05). Conclusion: RT-PCR with better applicability will become an efficient tool for detection of virulent Y. enterocolitica and will also be used as an alternative to the culture method. Due to the higher incidence than before, we should publicize dietary hygiene and pay more attention to earlier diagnosis for yersiniosis and its complications, aiming at children <4 years and adults >60 years during the autumn-winter months.

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“…Only automated bacterial blood culturing systems meet many of the requirements of an ideal test because they detect a wide range of organisms at concentrations of only 1 to 10 CFU per ml (5). Recently, molecular genetic techniques based on bacterial genomic detection were developed, with the 16S rRNA gene as a target (7,11,29,33). In real-time PCR, a sensitivity of about 30 CFU per ml was demonstrated (33).…”

mentioning

confidence: 99%

Two Novel Real-Time Reverse Transcriptase PCR Assays for Rapid Detection of Bacterial Contamination in Platelet Concentrates

2004

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The incidence of platelet bacterial contamination is approximately 1 per 2,000 units and has been acknowledged as the most frequent infectious risk from transfusion. In preliminary studies, the sterility of platelet concentrates (PCs) was tested with an automated bacterial blood culturing system and molecular genetic assays. Two real-time reverse transcriptase PCR (RT-PCR) assays performed in a LightCycler instrument were developed and compared regarding specificity and sensitivity by the use of different templates to detect the majority of the clinically important bacterial species in platelets. Primers and probes specific for the conserved regions of the eubacterial 23S rRNA gene or the groEL gene (encoding the 60-kDa heat shock protein Hsp60) were designed. During the development of the 23S rRNA RT-PCR, problems caused by the contamination of reagents with bacterial DNA were noted. Treatment with 8-methoxypsoralen and UV irradiation reduced the level of contaminating DNA. The sensitivity of the assays was greatly influenced by the enzyme system which was used. With rTth DNA polymerase in a one-enzyme system, we detected 500 CFU of Escherichia coli or Staphylococcus epidermidis/ml. With a two-enzyme system consisting of Moloney murine leukemia virus RT and Taq DNA polymerase, we detected 16 CFU/ml. With groEL mRNA as the target of RT-PCR under optimized conditions, we detected 125 CFU of E. coli/ml, and no problems with false-positive results caused by reagent contamination or a cross-reaction with human nucleic acids were found. Furthermore, the use of mRNA as an indicator of viability was demonstrated. Here we report the application of novel real-time RT-PCR assays for the detection of bacterial contamination of PCs that are appropriate for transfusion services.

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Direct identification of Yersinia enterocolitica in blood by polymerase chain reaction amplification

Cited by 67 publications

References 19 publications

Low Occurrence of Pathogenic Yersinia enterocolitica in Clinical, Food, and Environmental Samples: a Methodological Problem

Low Occurrence of Pathogenic Yersinia enterocolitica in Clinical, Food, and Environmental Samples: a Methodological Problem

Clinical Isolation and Characterization of Yersinia enterocolitica in China Using Real-Time PCR and Culture Method

Two Novel Real-Time Reverse Transcriptase PCR Assays for Rapid Detection of Bacterial Contamination in Platelet Concentrates

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