scholarly journals Identification of Neisseria gonorrhoeae isolates with a recombinant porA gene in Scotland, United Kingdom, 2010 to 2011

2012 ◽  
Vol 17 (9) ◽  
Author(s):  
K Eastick ◽  
A Winter ◽  
S Jamdar
Keyword(s):  
Polymerase Chain Reaction ◽  
United Kingdom ◽  
Neisseria Gonorrhoeae ◽  
Sequence Type ◽  
The Other ◽  
Chain Reaction ◽  
Polymerase Chain

Three isolates of Neisseria gonorrhoeae have been identified in Scotland in 2010 and 2011, which lack sequences in the porA pseudogene commonly used as the target for confirmatory gonorrhoea polymerase chain reaction assays. Two isolates were clustered temporally and geographically and have the same sequence type and porA sequence. A similar strain was reported in Australia during early 2011. The other Scottish isolate was identified separately and is different in sequence type and porA sequence.

scholarly journals Development of a Genoserotyping Method for Salmonella Infantis Detection on the Basis of Pangenome Analysis

2020 ◽  
Vol 9 (1) ◽  
pp. 67
Author(s):  
Seung-Min Yang ◽  
Jiwon Baek ◽  
Eiseul Kim ◽  
Hyeon-Be Kim ◽  
Seyoung Ko ◽  
...  
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Bacterial Species ◽  
Cost Effective ◽  
The Other ◽  
Gene Marker ◽  
Diagnostic Assay ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Substantial Economic Burden

In recent years, Salmonella Infantis has become a predominant serovariant in clinical and poultry isolates, thereby imposing a substantial economic burden on both public health and the livestock industry. With the aim of coping with the steep increase in serovar Infantis prevalence, a polymerase chain reaction (PCR)-based rapid and accurate diagnostic assay was developed in this study through pangenome profiling of 60 Salmonella serovars. A gene marker, SIN_02055, was identified, which is present in the S. Infantis genome but not in the pangenome of the other serovars. Primers specific to SIN_02055 were used to accurately detect serovar Infantis, and to successfully differentiate Infantis from the other 59 serovars in real-time PCR with a R2 of 0.999 and an efficiency of 95.76%. The developed method was applied to 54 Salmonella strains belonging to eight dominant serovars, and distinguished Infantis from the other seven serovars with an accuracy of 100%. The diagnostic primer set also did not show false positive amplification with 32 strains from eight non-Salmonella bacterial species. This cost-effective and rapid method can be considered an alternative to the classic serotyping using antisera.

scholarly journals Adult gonococcal keratoconjunctivitis: early detection is key

2020 ◽  
Vol 17 (1) ◽  
pp. 30-34
Author(s):  
Daniel Lai ◽  
Keith Ong
Keyword(s):  
Polymerase Chain Reaction ◽  
Early Detection ◽  
Neisseria Gonorrhoeae ◽  
Ophthalmic Solution ◽  
Careful Monitoring ◽  
Corneal Ulceration ◽  
Chain Reaction ◽  
Bacterial Conjunctivitis ◽  
Polymerase Chain ◽  
The Right

We describe a case of a 52-year-old male presenting with severe mucopurulent conjunctivitis of the right eye. Corneal ulceration and associated anterior chamber activity was noted later in the course of the disease. Neisseria gonorrhoeae was positive on polymerase chain reaction (PCR) testing earlier than traditional microscopy and culture. He was successfully treated with ceftriaxone 500 mg intravenously and azithromycin 1 g orally as single doses in addition to ofloxacin ophthalmic solution 0.3% hourly to the right eye. This case highlights the need to consider the possibility of gonococcus in cases of suspected bacterial conjunctivitis, careful monitoring for corneal involvement and the importance of early detection with PCR.

scholarly journals Detection of Neisseria gonorrhoeae in First-Voided Urine Sediments from Male Urethritis Patients by Polymerase Chain Reaction

1992 ◽  
Vol 66 (9) ◽  
pp. 1209-1212 ◽  
Author(s):  
Hisao KOMEDA ◽  
Takashi DEGUCHI ◽  
Hiroyuki YAMAMOTO ◽  
Hideki IWATA ◽  
Yasuhisa ITO ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Neisseria Gonorrhoeae ◽  
Chain Reaction ◽  
Polymerase Chain

Identification and diagnosis of Leishmania mexicana complex isolates by polymerase chain reaction

Parasitology ◽  
1994 ◽  
Vol 109 (4) ◽  
pp. 423-433 ◽  
Author(s):  
S. Eresh ◽  
S. M. McCallum ◽  
D. C. Barker
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
The Other ◽  
South American ◽  
Kinetoplast Dna ◽  
Leishmania Mexicana ◽  
Chain Reaction ◽  
Potential Host ◽  
Oligonucleotide Primers ◽  
Polymerase Chain

SUMMARYFollowing cloning of Leishmania (L.) amazonensis kinetoplast DNA two recombinant clones were identified: one specific for L. (L.) amazonensis and the other specific for L. (L.) amazonensis and closely related isolates. DNA sequences from these clones were compared with those of other kinetoplastids and oligonucleotide primers were designed to be used in the polymerase chain reaction. A pair of these primers has been shown not only to be highly specific for L. mexicana complex isolates but can also be used to distinguish between L. (L.) mexicana and L. (L.) amazonensis isolates. These primers have been tested with water-lysed cultures, crude DNA extracts from human patients, potential host reservoirs, sandfly vectors and with cell pellets after isoenzyme characterization. The results of these tests indicate that the primers can be used specifically in the presence of excess host DNA originating from the majority of South American countries.

Phenotypic and genotypic analysis of Flavobacterium psychrophilum isolates from Ontario salmonids with bacterial coldwater disease

2008 ◽  
Vol 54 (8) ◽  
pp. 619-629 ◽  
Author(s):  
Shohreh Hesami ◽  
Katie J. Allen ◽  
Devon Metcalf ◽  
Vaughn E. Ostland ◽  
Janet I. MacInnes ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Rainbow Trout ◽  
16S Rrna ◽  
Sequence Type ◽  
Chain Reaction ◽  
Flavobacterium Psychrophilum ◽  
Genotypic Analysis ◽  
Polymerase Chain ◽  
Coldwater Disease ◽  
Bacterial Coldwater Disease

Flavobacterium psychrophilum is the causative agent of bacterial coldwater disease (BCWD) and rainbow trout fry syndrome. BCWD has a considerable economic impact on aquaculture operations in Ontario, Canada, and our limited understanding of the population structure and epidemiology of F. psychrophilum isolates is an impediment to the development of improved management strategies. Seventy-five 16S rRNA gene and gyr polymerase chain reaction positive isolates of F. psychrophilum that had been collected over a 16-year period from farmed salmonids with tail rot, necrotic myositis, and osteochondrosis were characterized morphologically, biochemically, and genotypically. Although the isolates were homogeneous by preliminary biochemical and phenotypic characterization, two distinct biovars were found by API ZYM testing. As well, four restriction pattern types were detected by 16S rRNA polymerase chain reaction – restriction fragment length polymorphism analysis and there was a significant (P < 0.001) correlation between biovar I and digestion with MaeIII and between biovar II and digestion with MnlI or no site (P < 0.05). Further heterogenity was detected by sequence analysis of a 194 bp stem loop 3 region of rRNA. Nine sequence types were identified; 40/46 biovar I isolates were sequence type “a”, while 21/32 biovar II isolates belonged to either sequence type “c” or “d”. More than one biovar and genotype was identified among the strains recovered from separate fish sampled from three groups of rainbow trout ( Oncorhynchus mykiss ) experiencing BCWD mortality events. No association was found between genotype or biovar and type of disease. Taken together, these data suggest that F. psychrophilum from Ontario can be grouped into two major lineages based on biovar and 16S rRNA polymorphisms, and although three major strain types were most frequently isolated in this study, it appears that the population of F. psychrophilum with pathogenic potential is quite heterogeneous.

Verification of clinical samples, positive in AMPLICOR Neisseria gonorrhoeae polymerase chain reaction, by 16S rRNA and gyrA compared with culture

2005 ◽  
Vol 16 (6) ◽  
pp. 415-419 ◽  
Author(s):  
Åsa Airell ◽  
Emma Lindbäck ◽  
Ferda Ataker ◽  
Kirsti Jalakas Pörnull ◽  
Bengt Wretlind
Keyword(s):  
Polymerase Chain Reaction ◽  
16S Rrna ◽  
Neisseria Gonorrhoeae ◽  
False Negative ◽  
Clinical Samples ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Gyra Gene ◽  
Two Samples ◽  
Polymerase Chain

We compared 956 samples for AMPLICOR Neisseria gonorrhoeae polymerase chain reaction (PCR) (Roche) with species verification using the 16S rRNA gene to verification using gyrA gene. Control was the culture method. The gyrA verification uses pyrosequencing of the quinolone resistance-determining region of gyrA. Of 52 samples with optical density ≥0.2 in PCR, 27 were negative in culture, two samples from pharynx were false negative in culture and four samples from pharynx were false positives in verification with 16S rRNA. Twenty-five samples showed growth of gonococci, 18 of the corresponding PCR samples were verified by both methods; three urine samples were positive only in gyrA ; and one pharynx specimen was positive only in 16S rRNA. Three samples were lost. We conclude that AMPLICOR N. gonorrhoeae PCR with verification in gyrA gene can be considered as a diagnostic tool in populations with low prevalence of gonorrhoea and that pharynx specimens should not be analysed by PCR.

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