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

Sakai T, Kohzaki K, Watanabe A, Tsuneoka H, Shimadzu M - Clin Ophthalmol (2012)

Bottom Line: Thirteen samples of vitreous fluid (VF) were obtained from 13 patients during vitrectomy.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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Jikei University School of Medicine, Tokyo, Japan.

ABSTRACT

Background: To examine the utility of DNA microarray analysis for identifying causative microorganisms in endophthalmitis.

Methods: Thirteen 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.

Results: No 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.

Conclusions: Microarray 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.

No MeSH data available.


Related in: MedlinePlus

Results of the hybridization assay for polymerase chain reaction (PCR) amplifications from vitreous sample of case 1. PCR products are spotted as a reference on the right side. Green or blue (arrow) circles in V1, V2, and V3 represent strong hybridization. Bacterial identification is determined by the combination of strong hybridization in V1, V2, and V3. This case shows a pattern of Klebsiella pneumonia.
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f2-opth-6-321: Results of the hybridization assay for polymerase chain reaction (PCR) amplifications from vitreous sample of case 1. PCR products are spotted as a reference on the right side. Green or blue (arrow) circles in V1, V2, and V3 represent strong hybridization. Bacterial identification is determined by the combination of strong hybridization in V1, V2, and V3. This case shows a pattern of Klebsiella pneumonia.

Mentions: No control sample was positive for bacteria or fungi in microscopy, culture tests, PCR, or microarray analysis. The samples from Cases 1 and 2 were positive for bacteria in PCR analysis of VF specimens, and the sample from Case 3 was positive for fungus in PCR analysis. The DNA microarray was used to detect bacterial and fungal pathogens from positive PCR specimens. Data analysis revealed increased expression levels of genes from specific microorganisms in the VF samples. DNA microarray analysis identified Klebsiella pneumonia in Case 1 (Figure 2), Streptococcus agalactiae in Case 2 (Figure 3), and Candida parapsilosis in Case 3 (Figure 4) in the PCR-positive specimens. The results of the culture tests required 3 to 4 days to obtain, and were similarly positive for K. pneumonia in Case 1, S. agalactiae in Case 2, and C. parapsilosis in Case 3. The following paragraphs provide some background on each case.


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

Sakai T, Kohzaki K, Watanabe A, Tsuneoka H, Shimadzu M - Clin Ophthalmol (2012)

Results of the hybridization assay for polymerase chain reaction (PCR) amplifications from vitreous sample of case 1. PCR products are spotted as a reference on the right side. Green or blue (arrow) circles in V1, V2, and V3 represent strong hybridization. Bacterial identification is determined by the combination of strong hybridization in V1, V2, and V3. This case shows a pattern of Klebsiella pneumonia.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3295629&req=5

f2-opth-6-321: Results of the hybridization assay for polymerase chain reaction (PCR) amplifications from vitreous sample of case 1. PCR products are spotted as a reference on the right side. Green or blue (arrow) circles in V1, V2, and V3 represent strong hybridization. Bacterial identification is determined by the combination of strong hybridization in V1, V2, and V3. This case shows a pattern of Klebsiella pneumonia.
Mentions: No control sample was positive for bacteria or fungi in microscopy, culture tests, PCR, or microarray analysis. The samples from Cases 1 and 2 were positive for bacteria in PCR analysis of VF specimens, and the sample from Case 3 was positive for fungus in PCR analysis. The DNA microarray was used to detect bacterial and fungal pathogens from positive PCR specimens. Data analysis revealed increased expression levels of genes from specific microorganisms in the VF samples. DNA microarray analysis identified Klebsiella pneumonia in Case 1 (Figure 2), Streptococcus agalactiae in Case 2 (Figure 3), and Candida parapsilosis in Case 3 (Figure 4) in the PCR-positive specimens. The results of the culture tests required 3 to 4 days to obtain, and were similarly positive for K. pneumonia in Case 1, S. agalactiae in Case 2, and C. parapsilosis in Case 3. The following paragraphs provide some background on each case.

Bottom Line: Thirteen samples of vitreous fluid (VF) were obtained from 13 patients during vitrectomy.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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Jikei University School of Medicine, Tokyo, Japan.

ABSTRACT

Background: To examine the utility of DNA microarray analysis for identifying causative microorganisms in endophthalmitis.

Methods: Thirteen 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.

Results: No 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.

Conclusions: Microarray 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.

No MeSH data available.


Related in: MedlinePlus