scholarly journals Molecular Diagnosis of Periprosthetic Joint Infection by Quantitative RT-PCR of Bacterial 16S Ribosomal RNA

2013 ◽  
Vol 2013 ◽  
pp. 1-4 ◽  
Author(s):  
Mel S. Lee ◽  
Wen-Hsin Chang ◽  
Su-Chin Chen ◽  
Pang-Hsin Hsieh ◽  
Hsin-Nung Shih ◽  
...  
Keyword(s):  
16S Rrna ◽  
Molecular Diagnosis ◽  
Ribosomal Rna ◽  
Periprosthetic Joint Infection ◽  
Joint Infection ◽  
16S Ribosomal Rna ◽  
Purulent Discharge ◽  
Fistula Tract ◽  
Culture Negative

The diagnosis of periprosthetic joint infection is sometimes straightforward with purulent discharge from the fistula tract communicating to the joint prosthesis. However it is often difficult to differentiate septic from aseptic loosening of prosthesis because of the high culture-negative rates in conventional microbiologic culture. This study used quantitative reverse transcription polymerase chain reaction (RT-qPCR) to amplify bacterial 16S ribosomal RNA in vitro and in 11 clinical samples. The in vitro analysis demonstrated that the RT-qPCR method was highly sensitive with the detection limit of bacterial 16S rRNA being 0.148 pg/μl. Clinical specimens were analyzed using the same protocol. The RT-qPCR was positive for bacterial detection in 8 culture-positive cases (including aerobic, anaerobic, and mycobacteria) and 2 culture-negative cases. It was negative in one case that the final diagnosis was confirmed without infection. The molecular diagnosis of bacterial infection using RT-qPCR to detect bacterial 16S rRNA around a prosthesis correlated well with the clinical findings. Based on the promising clinical results, we were attempting to differentiate bacterial species or drug-resistant strains by using species-specific primers and to detect the persistence of bacteria during the interim period before the second stage reimplantation in a larger scale of clinical subjects.

scholarly journals Evaluation of 16S, map1 and pCS20 probes for detection of Cowdria and Ehrlichia species

1999 ◽  
Vol 122 (2) ◽  
pp. 323-328 ◽  
Author(s):  
M. T. E. P. ALLSOPP ◽  
C. M. HATTINGH ◽  
S. W. VOGEL ◽  
B. A. ALLSOPP
Keyword(s):  
16S Rrna ◽  
Ribosomal Rna ◽  
Pcr Amplification ◽  
16S Ribosomal Rna ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Amblyomma Hebraeum ◽  
Ribosomal Rna Gene ◽  
Group Iii ◽  
16S Ribosomal Rna Gene

A panel of 16S ribosomal RNA gene probes has been developed for the study of the epidemiology of heartwater; five of these detect different cowdria genotypes, one detects five distinct genotypes; one detects any Group III Ehrlichia species other than Cowdria and one detects any Group II Ehrlichia species. These probes have been used on PCR-amplified rickettsial 16S rRNA genes from over 200 Amblyomma hebraeum ticks. Control ticks were laboratory-reared and either uninfected or fed on sheep experimentally infected with different cowdria isolates, field ticks were collected from animals in heartwater-endemic areas. All tick-derived DNA samples were also examined by PCR amplification and probing for two other cowdria genes (map1 and pCS20) which have previously been used for heartwater epidemiology. This paper describes the first direct comparison of all currently available DNA probes for heartwater-associated organisms.

scholarly journals Detection of Nocardia by 16S Ribosomal RNA Gene PCR and Metagenomic Next-Generation Sequencing (mNGS)

2022 ◽  
Vol 11 ◽  
Author(s):  
Juanjuan Ding ◽  
Bing Ma ◽  
Xupeng Wei ◽  
Ying Li
Keyword(s):  
16S Rrna ◽  
Ribosomal Rna ◽  
Human Herpesvirus ◽  
Antibiotic Sensitivity ◽  
Common Species ◽  
16S Ribosomal Rna ◽  
Clinical Samples ◽  
Klebsiella Pneumonia ◽  
Antimicrobial Drugs ◽  
Detection And Identification

In this study, the aim was to investigate the discriminatory power of molecular diagnostics based on mNGS and traditional 16S ribosomal RNA PCR among Nocardia species. A total of fourteen clinical isolates from patients with positive Nocardia cultures and clinical evidence were included between January 2017 and June 2020 in HeNan Provincial People’s Hospital. DNA extraction and 16S rRNA PCR were performed on positive cultures, and pathogens were detected by mNGS in these same samples directly. Among the 14 Nocardia isolates, four species were identified, and N. cyriacigeorgica (8 cases) is the most common species. Twelve of the 14 Nocardia spp. isolates were identified by the two methods, while two strains of N. cyriacigeorgica were not identified by mNGS. All tested isolates showed susceptibility to trimethoprim-sulfamethoxazole (SXT), amikacin and linezolid. Apart from Nocardia species, other pathogens such as Acinetobacter baumannii, Klebsiella pneumonia, Aspergillus, Enterococcus faecalis, Human herpesvirus, etc., were detected from the same clinical samples by mNGS. However, these different pathogens were considered as colonization or contamination. We found that it is essential to accurately identify species for determining antibiotic sensitivity and, consequently, choosing antibiotic treatment. 16S rRNA PCR was useful for identification of nocardial infection among species, while this technique needs the clinicians to make the pre-considerations of nocardiosis. However, mNGS may be a putative tool for rapid and accurate detection and identification of Nocardia, beneficial for applications of antimicrobial drugs and timely adjustments of medication.

Phage Activity Against Planktonic and Biofilm Staphylococcus aureus Periprosthetic Joint Infection Isolates

2021 ◽  
Author(s):  
Katherine M. Caflisch ◽  
Robin Patel
Keyword(s):  
Staphylococcus Aureus ◽  
Successful Treatment ◽  
Periprosthetic Joint Infection ◽  
Joint Infection ◽  
Bacterial Isolates ◽  
Small Colony Variant ◽  
Small Colony ◽  
Planktonic State ◽  
Phage Activity

We recently reported the successful treatment of a case of periprosthetic joint infection (PJI) with phage. Phage activity against bacteria causing PJI has not been systematically evaluated. Here we examined the in vitro activity of seven lytic phages against 122 clinical isolates of Staphylococcus aureus recovered between April 1999 and February 2018 from subjects with PJI. Phages were assessed against planktonic and biofilm phenotypes. Activity of individual phages was demonstrated against up to 73% of bacterial isolates in the planktonic state and up to 100% of biofilms formed by isolates that were planktonically phage-susceptible. Susceptibility to phage was not correlated with small colony variant status. These results demonstrate that phages can infect S. aureus causing PJI in both planktonic and biofilm phenotypes, and thus are worthy of investigation as an alternative or addition to antibiotics in this setting.

Culture-negative periprosthetic joint infection: prevalence, aetiology, evaluation, recommendations, and treatment

2020 ◽  
Vol 44 (7) ◽  
pp. 1255-1261
Author(s):  
Irene Kalbian ◽  
Jung Wee Park ◽  
Karan Goswami ◽  
Young-Kyun Lee ◽  
Javad Parvizi ◽  
...  
Keyword(s):  
Periprosthetic Joint Infection ◽  
Joint Infection ◽  
Infection Prevalence ◽  
Culture Negative

Culture-Negative Periprosthetic Joint Infection

2014 ◽  
Vol 96 (5) ◽  
pp. 430-436 ◽  
Author(s):  
Javad Parvizi ◽  
Omer Faruk Erkocak ◽  
Craig J Della Valle
Keyword(s):  
Periprosthetic Joint Infection ◽  
Joint Infection ◽  
Culture Negative

Heterogeneity between 16S ribosomal RNA gene copies borne by one Desulfitobacterium strain is caused by different 100-200 bp insertions in the 5´ region

2007 ◽  
Vol 53 (1) ◽  
pp. 116-128 ◽  
Author(s):  
Richard Villemur ◽  
Philippe Constant ◽  
Annie Gauthier ◽  
Martine Shareck ◽  
Réjean Beaudet
Keyword(s):  
In Situ Hybridization ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Ribosomal Rna ◽  
16S Ribosomal Rna ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Gene Copies

Strains of Desulfitobacterium hafniense, such as strains PCP-1, DP7, TCE1, and TCP-A, have unusual long 16S ribosomal RNA (rRNA) genes due to an insertion of approximately 100 bp in the 5' region. In this report, we analyzed the 16S rRNA genes of different Desulfitobacterium strains to determine if such an insertion is a common feature of desulfitobacteria. We amplified this region by polymerase chain reaction (PCR) from eight Desulfitobacterium strains (D. hafniense strains PCP-1, DP7, TCP-A, TCE1, and DCB-2; D. dehalogenans; D. chlororespirans; and Desulfitobacterium sp. PCE1) and resolved each PCR product by denaturing gradient gel electrophoresis (DGGE). All strains had from two to seven DGGE- migrating bands, suggesting heterogeneity in their 16S rRNA gene copies. For each strain, the 5' region of the 16S rRNA genes was amplified and a clone library was derived. Clones corresponding to most PCR–DGGE migration bands were isolated. Sequencing of representative clones revealed that the heterogeneity was generated by insertions of 100–200 bp. An insertion was found in at least one copy of the 16S rRNA gene in all examined strains. In total, we found eight different types of insertions (INS1–INS8) that varied from 123 to 193 nt in length. Two-dimensional structural analyses of transcribed sequences predicted that all insertions would form an energetically stable loop. Reverse transcriptase – PCR experiments revealed that most of the observed insertions in the Desulfitobacterium strains were excised from the mature 16S rRNA transcripts. Insertions were not commonly found in bacterial 16S rRNA genes, and having a different insertion in several 16S rRNA gene copies borne by a single bacterial species was rarely observed. The function of these insertions is not known, but their occurrence can have an important impact in deriving 16S rRNA oligonucleotidic fluorescence in situ hybridization probes, as these insertions can be excised from 16S rRNA transcripts.Key words: Desulfitobacterium, 16S ribosomal RNA genes, heterogeneity, gene insertions, fluorescence in situ hybridization.

In vitro synthesis of 16S ribosomal RNA containing single base changes and assembly into a functional 30S ribosome

Biochemistry ◽  
1987 ◽  
Vol 26 (8) ◽  
pp. 2353-2364 ◽  
Author(s):  
W. Krzyzosiak ◽  
R. Denman ◽  
K. Nurse ◽  
W. Hellmann ◽  
M. Boublik ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
16S Ribosomal Rna ◽  
In Vitro Synthesis ◽  
Single Base

scholarly journals Molecular Identification of Bacterial Pathogen Infecting Coconut Leaf Beetle

Buletin Palma ◽  
2016 ◽  
Vol 16 (2) ◽  
pp. 147
Author(s):  
JELFINA C. ALOUW ◽  
DIANA NOVIANTI ◽  
MELDY L.A. HOSANG
Keyword(s):  
Polymerase Chain Reaction ◽  
16S Rrna ◽  
Serratia Marcescens ◽  
Molecular Identification ◽  
Ribosomal Rna ◽  
Causal Agent ◽  
16S Ribosomal Rna ◽  
Font Size ◽  
Chain Reaction ◽  
Polymerase Chain

<p><span style="font-size: medium;">ABSTRACT </span></p><p>Many species of microorganisms can cause diseases and mortality of insect pests. Accurate detection and identification of the entomophatogens are essential for development of biological control agent to the pest. Brontispa longissima, a serious and invasive pest of coconut, was infected by bacterium causing mortality of the larvae and pupae in coconut field. Objective of the research was to identify bacterium as a causal agent of the field-infected B. longissima using molecular  technique.  Research  was  conducted  between  April  and  August 2011.  Molecular  identification  using polymerase chain reaction (PCR) amplification of 16s ribosomal RNA of the infected larvae and sequencing of the gene showed that Serratia marcescens is the causal agent of the disease.</p><p>Keywords: Brontispa longissima, coconut, 16s rRNA, Serratia marcescens.</p><p> </p><p><span style="font-size: medium;">Identifikasi Molekular Bakteri Pathogen yang Menginfeksi Hama Daun Kelapa <br />Brontispa longissima(Coleoptera:Chrysomelidae)</span></p><p><span style="font-size: medium;">ABSTRAK </span></p><p>Banyak mikroorganisme dapat menimbulkan penyakit pada serangga hama.  Deteksi dan identifikasi yang akurat dari  pathogen  penyebab  penyakit  pada  serangga (entomopathogen)  hama  merupakan  tahap  yang  penting  dalam  pengembangan pengendalian biologi untuk hama tersebut.  Brontispa longissima sebagai hama penting dan bersifat  invasif pada tanaman kelapa diinfeksi oleh sejenis bakteri yang menyebabkan kematian larva dan pupa dari serangga  tersebut di lapangan. Penelitian ini bertujuan untuk mengidentifikasi organisme penyebab penyakit pada hama B. longissima dengan menggunakan teknik molekuler. Penelitian dilaksanakan pada bulan April sampai dengan Agustus  2011. Identifikasi bakteri dilakukan dengan mengamplifikasi 16s ribosomal RNA dari larva yang terinfeksi dengan menggunakan PCR (polymerase chain reaction).  Hasil analisis sekuens nukleotida 16s ribosomal RNA dari larva yang terinfeksi menunjukkan bahwa Serratia marcescens adalah bakteri penyebab dari penyakit tersebut.</p><p>Kata kunci: Brontispa longissima, kelapa, 16s rRNA, Serratia marcescens.</p>

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