PCR Detection and Identification of Bacterial Contaminants in Ocular Samples from Post- Operative Endophthalmitis

Abrishami, Majid; Hashemi, Behnam; Abrishami, Mojtaba; Abnous, Khalil; Razavi-Azarkhiavi, Kamal; Behravan, Javad

doi:10.7860/jcdr/2015/10291.5733

Cited by 8 publications

(5 citation statements)

References 27 publications

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“…The reported bacterial detection by eubacterial PCR assay from vitreous specimens is 56.4% (18/32 patients) from Iran,20 64% from France,16 75% (30/40) from Coimbatore, India,13 and 75.8% (44/58) from Chennai, India,21 whereas we could detect bacteria in 65.13% (127/195) by broad-range PCR assay versus 30.76% specimens by culture.…”

Section: Discussioncontrasting

confidence: 53%

Utility of broad-range 16S rRNA PCR assay versus conventional methods for laboratory diagnosis of bacterial endophthalmitis in a tertiary care hospital

et al. 2018

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BackgroundEndophthalmitis, a sight-threatening intraocular infection, can be of postsurgical, post-traumatic or endogenous origin. Laboratory diagnosis-based appropriate therapy can be vision-saving. Conventional culture-based laboratory diagnosis takes time and lacks sensitivity. In this study a broad-range PCR assay was assessed against conventional and automated culture methods in vitreous specimens for accurate microbiological diagnosis.AimsTo use broad-range PCR assay targeting 16S ribosomal RNA (rRNA) region of bacteria and to assess its performance vis-à-vis conventional and automated culture methods in the laboratory diagnosis of endophthalmitis.MethodsVitreous specimens from 195 patients with clinically diagnosed endophthalmitis were processed for classical and automated culture methods, antimicrobial sensitivity and broad-range PCR assay targeting 762 bp region of 16S rRNA followed by nucleotide sequencing by Sanger’s method. Causative agents were identified from the nucleotide sequences analysed against the GenBank database, and organisms were identified using the Clinical and Laboratory Standards Institute (CLSI) MM18A guidelines.ResultsBacteria could be detected from 127 (65.13%) of the 195 vitreous specimens by broad-range PCR assay; bacterial isolation was possible from 17 (8.7%) and 60 (30.76%) of these specimens by conventional and automated culture methods, respectively (p<0.0001). PCR assay could detect two uncultured bacterium, and in five cases the bacterial identity could not be determined from NCBI database matching.ConclusionBroad-range PCR assay could provide definitive microbial diagnosis within 24 hours in significantly more patients (p<0.0001). Some rare organisms could be detected, useful in treatment modalities. Automated culture was significantly more sensitive than conventional culture.

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Section: Discussioncontrasting

confidence: 53%

Utility of broad-range 16S rRNA PCR assay versus conventional methods for laboratory diagnosis of bacterial endophthalmitis in a tertiary care hospital

et al. 2018

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“…The other species are generally perceived of little clinical significance and they are commonly considered as contaminants in clinical cultures. However, recent reports indicate that these organisms can be responsible for local or systemic infections in humans [ 4 – 11 ].…”

Section: Introductionmentioning

confidence: 99%

Identification and Pathogenic Potential of Clinical Bacillus and Paenibacillus Isolates

et al. 2016

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The soil-related Bacillus and Paenibacillus species have increasingly been implicated in various human diseases. Nevertheless, their identification still poses problems in the clinical microbiology laboratory and, with the exception of Bacillus anthracis and Bacillus cereus, little is known on their pathogenicity for humans. In this study, we evaluated the use of matrix-assisted laser desorption—ionization time of flight mass spectrometry (MALDI-TOF MS) in the identification of clinical isolates of these genera and conducted genotypic and phenotypic analyses to highlight specific virulence properties. Seventy-five clinical isolates were subjected to biochemical and MALDI-TOF MS identification. 16S rDNA sequencing and supplemental tests were used to solve any discrepancies or failures in the identification results. MALDI-TOF MS significantly outperformed classical biochemical testing for correct species identification and no misidentification was obtained. One third of the collected strains belonged to the B. cereus species, but also Bacillus pumilus and Bacillus subtilis were isolated at high rate. Antimicrobial susceptibility testing showed that all the B. cereus, B. licheniformis, B. simplex, B. mycoides, Paenibacillus glucanolyticus and Paenibacillus lautus isolates are resistant to penicillin. The evaluation of toxin/enzyme secretion, toxin-encoding genes, motility, and biofilm formation revealed that B. cereus displays the highest virulence potential. However, although generally considered nonpathogenic, most of the other species were shown to swim, swarm, produce biofilms, and secrete proteases that can have a role in bacterial virulence. In conclusion, MALDI-TOF MS appears useful for fast and accurate identification of Bacillus and Paenibacillus strains whose virulence properties make them of increasing clinical relevance.

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“…Little is known about the other species of the genus that are generally considered as contaminant when found in clinical cultures and their potential pathogenicity has been scarcely investigated (Celandroni et al 2016 ). However, some reports indicate that these organisms can cause nosocomial infections or severe illness both in immunocompromised and immunocompetent humans, even if rarely (Idelevich et al 2013 ; Abrishami et al 2015 ).…”

Section: Introductionmentioning

confidence: 99%

Pathogenic potential of bacteria isolated from commercial biostimulants

et al. 2022

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Microbial-based products are a promising alternative to agrochemicals in sustainable agriculture. However, little is known about their impact on human health even if some of them, i.e., Bacillus and Paenibacillus species, have been increasingly implicated in different human diseases. In this study, 18 bacteria were isolated from 2 commercial biostimulants, and they were genotypically and phenotypically characterized to highlight specific virulence properties. Some isolated bacteria were identified as belonging to the genus Bacillus by BLAST and RDP analyses, a genus in-depth studied for plant growth-promoting ability. Moreover, 16S rRNA phylogenetic analysis showed that seven isolates grouped with Bacillus species while two and four clustered, respectively, with Neobacillus and Peribacillus. Unusually, bacterial strains belonging to Franconibacter and Stenotrophomonas were isolated from biostimulants. Although Bacillus species are generally considered nonpathogenic, most of the species have shown to swim, swarm, and produced biofilms, that can be related to bacterial virulence. The evaluation of toxins encoding genes revealed that five isolates had the potential ability to produce the enterotoxin T. In conclusion, the pathogenic potential of microorganisms included in commercial products should be deeply verified, in our opinion. The approach proposed in this study could help in this crucial step.

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PCR Detection and Identification of Bacterial Contaminants in Ocular Samples from Post- Operative Endophthalmitis

Cited by 8 publications

References 27 publications

Utility of broad-range 16S rRNA PCR assay versus conventional methods for laboratory diagnosis of bacterial endophthalmitis in a tertiary care hospital

Utility of broad-range 16S rRNA PCR assay versus conventional methods for laboratory diagnosis of bacterial endophthalmitis in a tertiary care hospital

Identification and Pathogenic Potential of Clinical Bacillus and Paenibacillus Isolates

Pathogenic potential of bacteria isolated from commercial biostimulants

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