Real-Time Polymerase Chain Reaction Detection of Methicillin-ResistantStaphylococcus aureus:Impact on Nosocomial Transmission and Costs

2007 ◽  
Vol 28 (10) ◽  
pp. 1134-1141 ◽  
Author(s):  
L. O. Conterno ◽  
J. Shymanski ◽  
K. Ramotar ◽  
B. Toye ◽  
C. van Walraven ◽  
...  

Objectives.To assess the impact of real-time polymerase chain reaction (PCR) detection of methicillin-resistantStaphylococcus aureus(MRSA) on nosocomial transmission and costs.Design.Monthly MRSA detection rates were measured from April 1, 2000, through December 31, 2005. Time series analysis was used to identify changes in MRSA detection rates, and decision analysis was used to compare the costs of detection by PCR and by culture.Setting.A 1,200-bed, tertiary care hospital in Canada.Patients.Admitted patients at high risk for MRSA colonization. MRSA detection using culture-based screening was compared with a commercial PCR assay.Results.The mean monthly incidence of nosocomial MRSA colonization or infection was 0.37 cases per 1,000 patient-days. The time-series model indicated an insignificant decrease of 0.14 cases per 1,000 patient-days per month (95% confidence interval, —0.18 to 0.46) after the introduction of PCR detection (P= .39). The mean interval from a reported positive result until contact precautions were initiated decreased from 3.8 to 1.6 days (P<.001). However, the cost of MRSA control increased from Can$605,034 to Can$771,609. Of 290 PCR-positive patients, 120 (41.4%) were placed under contact precautions unnecessarily because of low specificity of the PCR assay used in the study; these patients contributed 37% of the increased cost. The modeling study predicted that the cost per patient would be higher with detection by PCR (Can$96) than by culture (Can$67).Conclusion.Detection of MRSA by the PCR assay evaluated in this study was more costly than detection by culture for reducing MRSA transmission in our hospital. The cost benefit of screening by PCR varies according to incidences of MRSA colonization and infection, the predictive values of the assay used, and rates of compliance with infection control measures.

1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.


2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Yang Zhang ◽  
Chunyang Dai ◽  
Huiyan Wang ◽  
Yong Gao ◽  
Tuantuan Li ◽  
...  

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.


1994 ◽  
Vol 66 (1) ◽  
pp. 69-75 ◽  
Author(s):  
Toshimichi Yamamoto ◽  
Keiji Tamaki ◽  
Toshinori Kojima ◽  
Rieko Uchihi ◽  
Yoshinao Katsumata ◽  
...  

2016 ◽  
Vol 4 (2) ◽  
pp. 264-270 ◽  
Author(s):  
Aml Soliman ◽  
Asmaa Abdel Aal ◽  
Reham Afify ◽  
Noha Ibrahim

AIM: Aim was to detect Brain and Acute Leukemia, Cytoplasmic (BAALC) and ETS-related gene (ERG) expression in patients with acute myeloid leukemia (AML) as well as to study their biologic and prognostic impact on the disease outcome and survival.PATIENTS AND METHODS: The current study was carried out on 44 patients with denovo acute myeloid leukemia, as well as 44 age and sex matched controls. The quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was performed for estimation of BAALC and ERG expression.RESULTS: The current study was carried out on 44 patients with denovo acute myeloid leukemia, as well as 44 age and sex matched controls. The quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was performed for estimation of BAALC and ERG expression. BAALC was expressed in 36 (81.82%) of AML cases versus 10 (22.72%) of the control group which was highly statistically significant (P < 0.001). While ERG was positive in 39(88.64%) of cases and 8(18.18 %) of controls and that was also highly statistically significant (P < 0.001).CONCLUSION: Further researches still needed to clarify the role of BAALC and ERG in the pathogenesis of leukemia and their importance as targets for treatment of AML.


1996 ◽  
Vol 8 (2) ◽  
pp. 181-185 ◽  
Author(s):  
Patricia K. Holyoake ◽  
Gary F. Jones ◽  
Peter R. Davies ◽  
Dennis L. Foss ◽  
Michael P. Murtaugh

A polymerase chain reaction (PCR) assay was used to confirm the presence of ileal symbiont (IS) intracellularis in 3 swine herds with a history of proliferative enteritis (PE). Two pooled fecal specimens, each comprising 5 individual stool samples, were collected from pen floors to screen for the presence of IS intracellularis and determine the age range of pigs shedding the organism. IS intracellularis was detected in the feces of clinically normal 10–25week-old grower/finisher pigs, indicating that this age range of pigs was the main source of infection for younger nursery pigs. Shedding continued without clinical disease when 10–100 g/ton of tylosin or 10 g/ton of chlortetracycline was added to the feed. PCR testing of pooled fecal samples can be used to identify groups of pigs affected with PE. The results of this study indicate that this PCR assay has the potential to accurately assess the IS intracellularis infection status of swine herds and the association of IS intracellular-is with PE and growth performance.


2017 ◽  
Vol 8 (2) ◽  
pp. 19-22
Author(s):  
Abu Naser Ibne Sattar ◽  
Sanjida Khondakar Setu ◽  
Ahmed Abu Saleh ◽  
Sharmeen Ahmed

The prevalence of Tuberculous meningitis remains largely underestimated due to nonspecific clinical manifestation at early stages. A total of 20 CSF samples were studied from clinically suspected cases of tuberculous meningitis. All the samples were processed for ZN staining, TB culture on LJ media and TB-PCR using IS6110 and TRC4 primers by standard protocols. Of the total 20 CSF samples, 12 cases were diagnosed as TB meningitis. Most of the tuberculous meningitis cases were found positive by PCR using TRC4 primers (83.33%) followed by IS6110 primer (66.67%) and culture on L-J media(8.33%). None were found positive by ZN staining. TB-PCR usingTRC4 primer showed higher positivity than using IS6110 in detecting tuberculous meningitis, since some strains of MTB may lack the IS6110 element in their genome. PCR assay using TRC4 primer is superior in diagnosing tuberculous meningitis. This study was aimed to evaluate the diagnostic potentials of CSF polymerase chain reaction (PCR) by using TRC4 and IS6110 primers. Further the results were also compared with culture on Lowenstein-Jensen (LJ) media and AFB smear by Zeihl-Nelson (ZN) staining.Bangladesh J Med Microbiol 2014; 08 (02): 19-22


Author(s):  
Wan Huang ◽  
Jisheng Zhang ◽  
Lingyi Zeng ◽  
Chengru Yang ◽  
Lining Yin ◽  
...  

BackgroundThis study aimed to determine the molecular characteristics of carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates in a hospital in western Chongqing, southwestern China.MethodsA total of 127 unique CRKP isolates were collected from the Yongchuan Hospital of Chongqing Medical University, identified using a VITEK-2 compact system, and subjected to microbroth dilution to determine the minimal inhibitory concentration. Enterobacteriaceae intergenic repeat consensus polymerase chain reaction and multilocus sequence typing were used to analyze the homology among the isolates. Genetic information, including resistance and virulence genes, was assessed using polymerase chain reaction. The genomic features of the CRKP carrying gene blaKPC-2 were detected using whole-genome sequencing.ResultsST11 was the dominant sequence type in the homology comparison. The resistance rate to ceftazidime-avibactam in children was much higher than that in adults as was the detection rate of the resistance gene blaNDM (p &lt; 0.0001). Virulence genes such as mrkD (97.6%), uge (96.9%), kpn (96.9%), and fim-H (84.3%) had high detection rates. IncF (57.5%) was the major replicon plasmid detected, and sequencing showed that the CRKP063 genome contained two plasmids. The plasmid carrying blaKPC-2, which mediates carbapenem resistance, was located on the 359,625 base pair plasmid IncFII, together with virulence factors, plasmid replication protein (rep B), stabilizing protein (par A), and type IV secretion system (T4SS) proteins that mediate plasmid conjugation transfer.ConclusionOur study aids in understanding the prevalence of CRKP in this hospital and the significant differences between children and adults, thus providing new ideas for clinical empirical use of antibiotics.


2013 ◽  
Vol 12 (2) ◽  
pp. 101
Author(s):  
Gh. K. A. Al-kuzaay ◽  
Q. H. Kshash

This study was conducted for exam 348 milk samples from (clinically mastitic and other healthy cows) in many areas in AL-Diwanyia province by using CMT and bacteriological testing , which appeared that (64.9%) as percentage of mastitis ( clinically 15.9% , subclinically 84.0% ) Streptococcus agalactiae mastitis 13.2% ( 26.6% clinically , 73.3 % subclinicaly) diagnose by PCR assay by using specific primer (16SrRNA). Streptococcus agalactiae (30 isolates) after classical methods applied for streptococcus agalactiae identification (86 isolates).


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