Broad-Range Bacterial and Fungal DNA Amplification on Vitreous Humor from Suspected Endophthalmitis Patients

Varghese, Betsy; Rodrigues, Camilla; Deshmukh, Minal; Natarajan, Sundaram; Kamdar, Paritosh; Mehta, Ajita

doi:10.1007/bf03256207

Cited by 17 publications

(12 citation statements)

References 24 publications

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“…Data have been published evaluating the application of the broad‐range 16S rRNA sequencing method in clinical settings (Table ) . Table A lists several publications demonstrating the utility of 16S rRNA polymerase chain reaction (PCR) method to identify organisms cultured from clinical specimens .…”

Section: Discussionmentioning

confidence: 99%

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Clinical impact of the use of 16S rRNA sequencing method for the identification of “difficult‐to‐identify” bacteria in immunocompromised hosts

Bharadwaj

Swaminathan

Salimnia

et al. 2011

Transplant Infectious Dis

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Molecular method of 16S rRNA sequencing is reported to be helpful in the accurate identification of organisms with ambiguous phenotypic profiles. We analyzed the use of 16S rRNA sequencing method to identify clinically significant, "difficult-to-identify" bacteria recovered from clinical specimens, and evaluated its role in patient management and consequent clinical outcome. Among the 172 "difficult-to-identify" bacteria recovered over a 4-year period, 140 were gram-positive cocci or gram-negative bacilli; identification by 16S rRNA did not play a role in the management of patients infected with these bacteria. From 32 patients, 33 "difficult-to-identify" gram-positive bacilli were identified; the organisms were mycobacteria, Nocardia, Tsukamurella, Rhodococcus, and Gordonia. In 24 patients for whom clinical data were available, results from the 16S rRNA sequencing method led to treatment change in 14 immunocompromised patients (including 7 hematopoietic stem cell recipients and 1 liver transplant recipient). Therapy was modified in 9 patients, initiated in 3 patients, and discontinued in 2 patients. Most patients' therapy was switched to oral antibiotics with discontinuation of intravascular catheters, facilitating early hospital discharge. All 14 patients were alive 30 days after infection onset. The present study demonstrates the clinical application of 16S rRNA sequencing method to identify "difficult-to-identify" mycobacteria and other gram-positive bacilli in clinical specimens, particularly in immunocompromised hosts.

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Section: Discussionmentioning

confidence: 99%

“…The 16S rRNA sequencing method has been documented to play an important role in the accurate identification of bacterial isolates, and in the discovery of novel bacteria in clinical microbiology laboratories. In addition, the novel method has been employed to identify bacteria directly from clinical specimens .…”

mentioning

confidence: 99%

Clinical impact of the use of 16S rRNA sequencing method for the identification of “difficult‐to‐identify” bacteria in immunocompromised hosts

Bharadwaj

Swaminathan

Salimnia

et al. 2011

Transplant Infectious Dis

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“…These include excised heart valves in suspected bacterial endocarditis (16,50,89,153), bloodstream infections in neonates or hematologyoncology patients (69,80,103,110,138), cerebrospinal fluid in suspected meningitis or shunt infections (32,73,134,158,167), bone samples or synovial fluid samples in osteomyelitis or septic arthritis (40, 41, 127), vitreous or aqueous fluid in intraocular infections (152), tissue or pus from brain or spinal abscesses (75,115), amniotic fluid in chorioamnionitis or preterm labor (34,58), and various other scenarios.…”

Section: Clinical Applicationsmentioning

confidence: 99%

Broad‐Range PCR for Detection and Identification of Bacteria

Maiwald¹

2011

Molecular Microbiology

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“…Rapid detection and identification of the causative pathogens is crucial for vision-saving diagnosis and treatment. Molecular techniques using multiplex or broad-range PCR enable rapid detection and identification of causative pathogens in ocular infectious diseases 1–3. Several problems remain to be solved, however.…”

Section: Introductionmentioning

confidence: 99%

Use of DNA microarray analysis in diagnosis of bacterial and fungal endophthalmitis

Sakai¹,

Kohzaki²,

Watanabe³

et al. 2012

OPTH

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BackgroundTo examine the utility of DNA microarray analysis for identifying causative microorganisms in endophthalmitis.MethodsThirteen samples of vitreous fluid (VF) were obtained from 13 patients during vitrectomy. Vitreous fluids from three patients with suspected endophthalmitis and ten controls without infection were subjected to testing for the presence of bacteria and fungi in culture tests, polymerase chain reaction (PCR) analysis, and DNA microarray analysis.ResultsNo control sample was positive for bacteria or fungi in the culture test, PCR, or microarray analysis. Specimens from two patients (Cases 1 and 2) with suspected endophthalmitis were positive for bacteria in PCR, and a specimen from one patient (Case 3) was positive for fungi in PCR. Klebsiella pneumonia (Case 1), Streptococcus agalactiae (Case 2), and Candida parapsilosis (Case 3) in the PCR-positive specimens were identified by DNA microarray analysis within 24 hours. Culture results were also positive for K. pneumonia in Case 1, S. agalactiae in Case 2, and C. parapsilosis in Case 3, but required 3 to 4 days to obtain.ConclusionsMicroarray analysis is complementary to routine cultures for identifying causative microorganisms and is likely to be a useful tool in patients with suspected endophthalmitis who require rapid diagnosis and early antibiotic treatment.

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Broad-Range Bacterial and Fungal DNA Amplification on Vitreous Humor from Suspected Endophthalmitis Patients

Cited by 17 publications

References 24 publications

Clinical impact of the use of 16S rRNA sequencing method for the identification of “difficult‐to‐identify” bacteria in immunocompromised hosts

Clinical impact of the use of 16S rRNA sequencing method for the identification of “difficult‐to‐identify” bacteria in immunocompromised hosts

Broad‐Range PCR for Detection and Identification of Bacteria

Use of DNA microarray analysis in diagnosis of bacterial and fungal endophthalmitis

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