scholarly journals Molecular Epidemiology of Rhodococcus equi Based on traA, vapA, and vapB Virulence Plasmid Markers

2007 ◽  
Vol 196 (5) ◽  
pp. 763-769 ◽  
Author(s):  
Alain A Ocampo-Sosa ◽  
Deborah A Lewis ◽  
Jesús Navas ◽  
Frances Quigley ◽  
Raquel Callejo ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Typing ◽  
Rhodococcus Equi ◽  
Virulence Plasmid ◽  
Chain Reaction ◽  
Gene Markers ◽  
Typing System ◽  
New Type ◽  
Polymerase Chain ◽  
Related Plasmid

AbstractMolecular typing of the actinomycete Rhodococcus equi is insufficiently developed, and little is known about the epidemiology and transmission of this multihost pathogen. We report a simple, reliable polymerase chain reaction typing system for R. equi based on 3 plasmid gene markers: traA from the conserved conjugal transfer machinery and vapA and vapB, found in 2 different plasmid subpopulations. This “TRAVAP” typing scheme classifies R. equi into 4 categories: traA+/vapA+B−, traA+/vapA−B+, traA+/vapAB−, and traA−/vapAB− (plasmidless). A TRAVAP survey of 215 R. equi strains confirmed the strong link between vapA (traA+/vapA+B− plasmids) and horse isolates and revealed other host-related plasmid associations: between traA+/vapA−B+ and pigs and between traA+/vapAB−—a new type of R. equi plasmid—and cattle. Plasmidless strains were more frequent among isolates from nonpathological specimens. All plasmid categories were common in human isolates, which possibly reflects the predominantly opportunistic nature of R. equi infection in this host and a zoonotic origin.

scholarly journals Rhodococcus equi Infections in Dogs

2016 ◽  
Vol 54 (1) ◽  
pp. 159-163 ◽  
Author(s):  
L. K. Bryan ◽  
S. D. Clark ◽  
J. Díaz-Delgado ◽  
S. D. Lawhon ◽  
J. F. Edwards
Keyword(s):  
At Risk ◽  
Polymerase Chain Reaction ◽  
Companion Animals ◽  
Immunosuppressive Drugs ◽  
Rhodococcus Equi ◽  
Virulence Plasmid ◽  
The Novel ◽  
Chain Reaction ◽  
Polymerase Chain

Five cases of Rhodococcus equi infection in dogs were identified from 2003 to 2014. Three of the dogs had severe, internal lesions attributable to R. equi that have not been previously described: endophthalmitis, endocarditis, and suppurative pleuropneumonia. Isolates from 4 of the dogs were analyzed by polymerase chain reaction for Rhodococcus virulence-associated plasmid (vap) genes. One isolate was vapA-positive, 2 lacked a virulence plasmid, and 1 carried the novel vapN-associated plasmid (pVAPN) recently characterized in bovine isolates. The pVAPN plasmid has not been described in isolates cultured from companion animals. Four of the dogs either were receiving immunosuppressive drugs or had endocrinopathies. R. equi has the potential to cause significant infections in dogs, and immunocompromised animals should be considered at risk for infection.

scholarly journals Rhodococcus equi Infections in Goats: Characterization of Virulence Plasmids

2017 ◽  
Vol 55 (2) ◽  
pp. 273-276 ◽  
Author(s):  
Lauren W. Stranahan ◽  
Quinci D. Plumlee ◽  
Sara D. Lawhon ◽  
Noah D. Cohen ◽  
Laura K. Bryan
Keyword(s):  
Polymerase Chain Reaction ◽  
Lymph Nodes ◽  
Rhodococcus Equi ◽  
Caseous Lymphadenitis ◽  
Chain Reaction ◽  
Disseminated Infection ◽  
Virulence Plasmids ◽  
Visceral Organs ◽  
Polymerase Chain

Rhodococcus equi is an uncommon cause of systemic pyogranulomatous infections in goats with macroscopic similarities to caseous lymphadenitis caused by Corynebacterium pseudotuberculosis. Caprine cases have previously been reported to be caused by avirulent R. equi strains. Six cases of R. equi infection in goats yielding 8 R. equi isolates were identified from 2000 to 2017. Lesions varied from bronchopneumonia, vertebral and humeral osteomyelitis, and subcutaneous abscesses, to disseminated infection involving the lungs, lymph nodes, and multiple visceral organs. Isolates of R. equi from infected goats were analyzed by polymerase chain reaction for R. equi virulence-associated plasmid ( vap) genes. Seven of 8 isolates carried the VapN plasmid, originally characterized in bovine isolates, while 1 isolate lacked virulence plasmids and was classified as avirulent. The VapN plasmid has not been described in isolates cultured from goats.

scholarly journals The Inc FII Plasmid and its Contribution in the Transmission of blaNDM-1 and blaKPC-2 in Klebsiella pneumoniae in Egypt

Antibiotics ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 266 ◽  
Author(s):  
Eman Ramadan Mohamed ◽  
Mamdouh Yones Ali ◽  
Nancy G F M Waly ◽  
Hamada Mohamed Halby ◽  
Rehab Mahmoud Abd El-Baky
Keyword(s):  
Polymerase Chain Reaction ◽  
Klebsiella Pneumoniae ◽  
Molecular Typing ◽  
University Hospital ◽  
Chain Reaction ◽  
Great Problem ◽  
E Coli ◽  
String Test ◽  
Resistance Patterns ◽  
Polymerase Chain

The emergence of blaKPC-2 and blaNDM-1 producing Klebsiella pneumoniae represents a great problem in many Egyptian hospitals. One hundred and twenty-six K. pneumoniae isolates from patients admitted to Assiut University Hospital were identified by an API20E kit. Carbapenemase-producing K. pneumoniae (CPKP) was detected by the modified carbapenem inactivation method (mCIM), the EDTA-modified carbapenem inactivation method (eCIM), and an E-test. Based on the polymerase chain reaction, all isolates were negative for bla-VIM-1 and bla-IMP-1, fifteen of these isolates were positive for both blaKPC-2 and blaNDM-1, two isolates were positive for blaKPC-2 only, and twenty-eight isolates were positive for bla-NDM-1 only. Although one isolate was positive for the string test, all CPKP isolates were negative for capsular genes. Only 71.1% of CPKP transferred their plasmids to their corresponding transconjugants (E. coli J53). The resistance patterns of the clinical isolates and their transconjugates were similar, except for 12 isolates, which showed differences with their transconjugates in the resistance profile of four antibiotics. Molecular typing of the plasmids based on replicon typing showed that Inc FIIK and FII plasmids predominated in isolates and their transconjugants carrying blaKPC-2 and/or blaNDM-1. Conjugative Inc FII plasmids play an important role in the spread of CPKP, and their recognition is essential to limit their spread.

Polymerase Chain Reaction-Based Detection of Circulating Melanoma Cells as an Effective Marker of Tumor Progression

1999 ◽  
Vol 17 (1) ◽  
pp. 304-304 ◽  
Author(s):  
Giuseppe Palmieri ◽  
Maria Strazzullo ◽  
Paolo A. Ascierto ◽  
Sabrina M.R. Satriano ◽  
Antonio Daponte ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Tumor Progression ◽  
Peripheral Blood ◽  
Melanoma Cells ◽  
Disease Stage ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Gene Markers ◽  
Risk Of Recurrence ◽  
Polymerase Chain

PURPOSE: Reverse transcriptase (RT) polymerase chain reaction (PCR) with multiple markers has been demonstrated to be highly sensitive in detecting circulating cells from patients with malignant melanoma (MM). We evaluated the clinical significance of the presence in peripheral blood of specific PCR-positive mRNA markers as an expression of circulating melanoma cells. PATIENTS AND METHODS: Total cellular RNA was obtained from the peripheral blood of 235 patients with either localized (n = 154) or metastatic (n = 81) melanoma. We performed RT-PCR using tyrosinase, p97, MUC18, and MelanA/MART1 as gene markers. The PCR products were analyzed by gel electrophoresis and Southern blot hybridization. In addition, 20 healthy subjects and 21 patients with nonmelanoma cancer were used as negative controls. RESULTS: Although detected at various levels among assessable patients, each mRNA marker was significantly correlated with disease stage. A significant correlation with disease stage was demonstrated for patients who were positive to all four markers (P < .0001) or to at least three markers (P < .001). Univariate analysis showed a significant correlation between risk of recurrence (evaluated in stage I, II, and III patients) and increasing number of PCR-positive markers (P = .0002). Logistic regression multivariate analysis indicated that each single marker (except tyrosinase) and, more especially, the presence of four PCR-positive markers remained statistically independent prognostic factors for tumor progression. CONCLUSION: Our data establish the existence of a significant correlation among clinical stages, tumor progression, and presence of circulating melanoma-associated antigens in peripheral blood of MM patients. Preliminary assessment of a subset of patients with a higher risk of recurrence needs longer follow-up and further studies to define the role of RT-PCR in monitoring MM patients.

Molecular Typing ofMycobacterium chelonaeIsolates From a Pseudo-outbreak Involving an Automated Bronchoscope Washer

2008 ◽  
Vol 29 (11) ◽  
pp. 1088-1090 ◽  
Author(s):  
Alexandra Chroneou ◽  
Sarah K. Zimmerman ◽  
Steven Cook ◽  
Sandra Willey ◽  
Jane Eyre-Kelly ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Bronchoalveolar Lavage ◽  
Molecular Typing ◽  
Bronchoalveolar Lavage Fluid ◽  
Lavage Fluid ◽  
Clinical Findings ◽  
Chain Reaction ◽  
Mycobacterium Chelonae ◽  
Polymerase Chain ◽  
Repetitive Extragenic Palindromic

We describe a pseudo-outbreak ofMycobacterium chelonaeinfection in bronchoalveolar lavage fluid from 9 patients that was traced to contamination of an automated bronchoscope washer. Molecular typing using repetitive extragenic palindromic polymerase chain reaction was helpful in confirming epidemiologic and clinical findings.

A quantitative reverse transcriptase polymerase chain reaction assay to identify metastatic carcinoma tissue of origin

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 20024-20024
Author(s):  
D. Talantov ◽  
J. Baden ◽  
T. Jatkoe ◽  
J. Yu ◽  
D. Atkins ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Primary Tumor ◽  
Metastatic Carcinoma ◽  
Unknown Primary ◽  
Chain Reaction ◽  
Gene Markers ◽  
Carcinoma Of Unknown Primary ◽  
Polymerase Chain ◽  
Tissue Of Origin

20024 Background: Carcinoma of unknown primary (CUP) wherein metastatic disease presents without an identifiable primary tumor site represents approximately 3–5% of all cancers. Identifying the origin of the primary tumor in patients with CUP can enable rational choice of therapeutic regimens. We developed an optimized set of ten gene markers for a quantitative reverse transcriptase polymerase chain reaction (qRTPCR) assay, and demonstrated high accuracy in predicting the tissue of origin when used on with formalin-fixed, paraffin-embedded (FFPE) metastatic carcinoma samples. Methods: Twenty-three putative tissue-specific markers for lung, colon, pancreas, breast, prostate and ovarian carcinomas were nominated by querying a gene expression profile database and by performing a literature search. Ten of these marker candidates were then selected based on validation by qRTPCR on 205 FFPE metastatic carcinomas of known tissue origin. Next, we optimized the RNA isolation and qRTPCR methods for these ten markers, and tested the qRTPCR assay on two sets of FFPE metastatic tumors. Results: We applied the 10-gene qRTPCR assay to a set of 260 metastatic tumors of known origin, generating an overall accuracy of 78%. Furthermore we tested an independent set of 48 metastatic samples, including thirty-seven samples where either the tissue of origin was known or which initially presented as CUP but were subsequently resolved. In these 48 samples, our assay demonstrated an accuracy of 76%. Conclusions: Our results suggested that optimized ten-gene markers qRTPCR assay reliably predicts tissue of origin of metastatic carcinomas in FFPE tissues. Such assay can significantly improve the rate of tissue of origin identification for carcinoma of unknown primary. [Table: see text]

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