Use of BACTEC radiometric culture method and polymerase chain reaction for the rapid screening of faeces and tissues for Mycobacterium paratuberculosis

1995 ◽  
Vol 72 (12) ◽  
pp. 458-462 ◽  
Author(s):  
DV COUSINS ◽  
RJ EVANS ◽  
BR FRANCIS
Keyword(s):  
Polymerase Chain Reaction ◽  
Culture Method ◽  
Rapid Screening ◽  
Mycobacterium Paratuberculosis ◽  
Chain Reaction ◽  
Polymerase Chain

Using a Polymerase Chain Reaction as a Complementary Test to Improve the Detection of Dermatophyte Fungus in Nails

2014 ◽  
Vol 104 (3) ◽  
pp. 233-237 ◽  
Author(s):  
María José Iglesias Sánchez ◽  
Ana María Pérez Pico ◽  
Félix Marcos Tejedor ◽  
María Jesús Iglesias Sánchez ◽  
Raquel Mayordomo Acevedo
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Classical Method ◽  
Clinical Manifestations ◽  
Culture Method ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Traditional Procedure ◽  
Polymerase Chain

Background Dermatomycoses are a group of pathologic abnormalities frequently seen in clinical practice, and their prevalence has increased in recent decades. Diagnostic confirmation of mycotic infection in nails is essential because there are several pathologic conditions with similar clinical manifestations. The classical method for confirming the presence of fungus in nail is microbiological culture and the identification of morphological structures by microscopy. Methods We devised a nested polymerase chain reaction (PCR) that amplifies specific DNA sequences of dermatophyte fungus that is notably faster than the 3 to 4 weeks that the traditional procedure takes. We compared this new technique and the conventional plate culture method in 225 nail samples. The results were subjected to statistical analysis. Results We found concordance in 78.2% of the samples analyzed by the two methods and increased sensitivity when simultaneously using the two methods to analyze clinical samples. Now we can confirm the presence of dermatophyte fungus in most of the positive samples in just 24 hours, and we have to wait for the result of culture only in negative PCR cases. Conclusions Although this PCR cannot, at present, substitute for the traditional culture method in the detection of dermatophyte infection of the nails, it can be used as a complementary technique because its main advantage lies in the significant reduction of time used for diagnosis, in addition to higher sensitivity.

scholarly journals Direct Polymerase Chain Reaction-Based Detection of Cercospora beticola in Field Soils

Plant Disease ◽  
2010 ◽  
Vol 94 (9) ◽  
pp. 1100-1104 ◽  
Author(s):  
R. T. Lartey ◽  
T. C. Caesar-TonThat ◽  
A. W. Lenssen ◽  
J. Eckhoff ◽  
S. L. Hanson ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sugar Beet ◽  
Fragment Size ◽  
Rapid Screening ◽  
Cercospora Beticola ◽  
Field Soil ◽  
Chain Reaction ◽  
Field Soils ◽  
Total Dna ◽  
Polymerase Chain

Cercospora beticola, the causal agent of Cercospora leaf spot of sugar beet, survives as pseudostromata in infected sugar beet residues in the soil. Under optimal conditions, overwintering propagules germinate and produce conidia that are dispersed as primary inoculum to initiate infection in sugar beet. We developed a polymerase chain reaction (PCR) technique for rapid detection of C. beticola in field soils. Total DNA was first isolated from soil amended with C. beticola culture using the PowerSoil DNA Kit. The purified DNA was subjected to PCR in Extract-N-Amp PCR mix with CBACTIN primers over 35 cycles. The amplified products were resolved and compared by electrophoresis in 1% agarose gels. The PCR fragment size of C. beticola from the amended field soil correlated in size with the amplicon from control C. beticola culture DNA extract. Additionally, sample soils were collected from sugar beet fields near Sidney, MT and Foxholm, ND. Total DNA was extracted from the samples and subjected to PCR and resolved as previously described. The amplicons were purified from the gels and subjected to BigDye Terminator Cycle sequencing. All sequences from field soils samples, C. beticola-amended field soil, and pure culture were compared by alignment with a C. beticola actin gene sequence from GenBank. The result of the alignment confirmed the amplicons as products from C. beticola. Rapid screening for the presence of C. beticola in the soil by PCR will improve research capabilities in biological control, disease forecasting, and management of this very important sugar beet pathogen.

Detection of legionella bacteria in sewage by polymerase chain reaction and standard culture method

1995 ◽  
Vol 31 (5-6) ◽  
pp. 409-416 ◽  
Author(s):  
Bruce M. Roll ◽  
Roger S. Fujioka
Keyword(s):  
Polymerase Chain Reaction ◽  
Sewage Treatment ◽  
Culture Method ◽  
Chain Reaction ◽  
Standard Culture ◽  
Pontiac Fever ◽  
Legionella Species ◽  
Legionnaires Disease ◽  
The Subject ◽  
Polymerase Chain

Legionella bacteria are ubiquitous in environmental waters. Only a few species of Legionella , especially, L. pneumophila are pathogenic to humans and cause a sometimes fatal Legionnaires disease as well as a less fatal disease called Pontiac fever. The presence of Legionella in sewage and aerosolized sewage is the subject of this investigation because reuse of sewage may involve the exposure of people to aerosolization, the mode of transmission of Legionella bacteria. The objective of this study was to determine the prevalence of Legionella species and L. pneumophila in wastewater and their fate after various stages of treatment. The polymerase chain reaction (PCR) and standard culture method were utilized to detect Legionella species and L. pneumophila. PCR results indicated that Legionella species were present at levels > 103 cells / ml during all phases of sewage treatment including chlorinated effluents. Culture results indicated levels at least one log lower than seen with PCR. Legionella species were also recovered from air samples collected from secondary aeration basins at levels < 103 cells/ml. PCR was shown to be the most rapid and sensitive method for detecting Legionella in sewage.

scholarly journals Search for Mycobacterium paratuberculosis DNA in orofacial granulomatosis and oral Crohn’s disease tissue by polymerase chain reaction

Gut ◽  
1997 ◽  
Vol 41 (5) ◽  
pp. 646-650 ◽  
Author(s):  
M P Riggio ◽  
J Gibson ◽  
A Lennon ◽  
D Wray ◽  
D G MacDonald
Keyword(s):  
Polymerase Chain Reaction ◽  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Oral Lesions ◽  
Mycobacterium Paratuberculosis ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Orofacial Granulomatosis ◽  
Polymerase Chain ◽  
Normal Controls

Background—Although intestinal Crohn’s disease has long been suspected to have a mycobacterial cause, possible mycobacterial involvement in orofacial granulomatosis (OFG) and oral lesions of Crohn’s disease has not yet been investigated.Aims—As the slow growingMycobacterium paratuberculosis has been implicated in the aetiology of intestinal Crohn’s disease, the potential involvement of this mycobacterial species in OFG and oral lesions of Crohn’s disease was investigated.Patients—To attempt detection of the organism in OFG and oral Crohn’s disease tissue samples, a polymerase chain reaction (PCR) assay was used on archival formalin fixed, paraffin wax embedded oral tissue sections from 30 patients with OFG, seven with Crohn’s disease, and 12 normal controls.Methods—The PCR assay used was based on primers targeting the 5′ region of the multicopy IS900 DNA insertion element of the M paratuberculosis genome. In order to achieve maximum sensitivity, two rounds of PCR were carried out and amplicons confirmed by Southern blot hybridisation to a digoxigenin labelled IS900 DNA probe.Results—None of the OFG and oral lesions of Crohn’s disease samples were positive forMparatuberculosis and all normal controls were also negative.Conclusions—These results suggest that M paratuberculosis may not be a major aetiological agent in OFG or oral Crohn’s disease lesions, although the use of paraffin wax embedded tissue as opposed to fresh tissue as a sample source could underestimate the true prevalence of the organism.

Detection of Mycobacterium paratuberculosis by Polymerase Chain Reaction in Children with Crohn's Disease

1994 ◽  
Vol 169 (2) ◽  
pp. 449-451 ◽  
Author(s):  
B. Dell'Isola ◽  
C. Poyart ◽  
O. Goulet ◽  
J. F. Mougenot ◽  
E. S. Journo ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Mycobacterium Paratuberculosis ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Detection of Campylobacter spp. in children diarrhea by using Polymerase Chain Reaction PCR technique in Al-Diwanyiah Governorate

2011 ◽  
Vol 10 (2) ◽  
pp. 45
Author(s):  
A. H. AL-Hamadani And Z.F.Saleh
Keyword(s):  
Polymerase Chain Reaction ◽  
Culture Method ◽  
Laboratory Diagnosis ◽  
Routine Laboratory ◽  
Direct Examination ◽  
Chain Reaction ◽  
Culture Methods ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Campylobacter Spp

This study was conducted in order to identify Campylobacter spp. As a causative agent of diarrhea in children using routine laboratory diagnosis (direct and culture methods) in comparison with polymerase chain reaction (PCR) technique as a confirm diagnostic tool. A total of 100 children stool samples were collected from both sexes at ages less than two years old suffering from diarrhea who admitted the maternity and Pediatric Teaching hospital in Al-Diwaniyiah Governorate from December 2007 to August 2008. Based on the clinical and laboratory diagnosis, results revealed that the percent of Campylobacter isolation was 8% included C. coli and C. jejuni for children samples. In addition, the results haven't revealed any statistically significant (P≥0.01) between the rate of infection and sexes, while there was a statistically significant (P≤0.01) between these rates and ages, where it noted that patients (>1) years old were more prone to infect with Campylobacter spp. exposure to infections. The results revealed that the PCR positive samples contained one band of amplified DNA with a molecular weight (816 bp) after electrophoresis and examined under UV- transilluminator. The study also showed that the sensitivity and specificity of PCR technique were 40% and 100% respectively for examination children samples, when compared with direct examination, but were with culture method were 33% and 100%; respectively in children.

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