Literature DB >> 24958188

Comparison of polymerase chain reaction and standard microbiological techniques in presumed bacterial corneal ulcers.

Anita Panda1, Tajinder Pal Singh, Geeta Satpathy, Meenakshi Wadhwani.   

Abstract

To assess utility of PCR in the diagnosis of bacterial corneal ulcer and to compare sensitivity and specificity of this technique with conventional laboratory methods. A prospective nonrandomized investigative study conducted on 122 eyes of presumed bacterial keratitis. Samples were collected for bacterial and fungal culture and Gram stain smear. A separate sample was taken for PCR with 26 gauge needle and was dipped directly into Eppendorff tube with lysis buffer in it. Diagnosis of culture proven bacterial keratitis was established in 53 (43.4 %) and most common bacterial isolate was staphylococcal sp. (83 %). Direct microscopic examination of Gram stained smear revealed presence of bacteria in 24 (23.9 %) specimens and PCR positivity was evident in 56 (45.5 %). In preantibiotic treated eyes culture was positive in 15 (30 %), Gram stain in 9 (18 %), and PCR in 18 (36 %). The same for untreated (fresh) eyes, positivity of culture as well as PCR was noted in 38 (52.7 %) and that of Gram stain was noted in 20 (27.7 %). Sensitivity of Gram stain and PCR was 45.28 and 88.68 % respectively; whereas specificity was 92.75 % for Gram stain and 86.96 % for PCR. The average time taken for PCR reaction was 4-8 h while culture reporting took at least 24-48 h. Our findings suggest that PCR is a good adjunct modality to the "Gold Standard" technique in the diagnosis of bacterial corneal ulcer.

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Year:  2014        PMID: 24958188     DOI: 10.1007/s10792-014-9925-9

Source DB:  PubMed          Journal:  Int Ophthalmol        ISSN: 0165-5701            Impact factor:   2.031


  29 in total

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Authors:  Visvaraja Subrayan; Mohammadreza Peyman; Soon Lek Yap; Nadir Ali Mohamed Ali; Shamala Devi
Journal:  Eye Contact Lens       Date:  2010-07       Impact factor: 2.018

2.  Corneal ulceration in the developing world--a silent epidemic.

Authors:  J P Whitcher; M Srinivasan
Journal:  Br J Ophthalmol       Date:  1997-08       Impact factor: 4.638

3.  Detection of resistance to isoniazid, rifampin, and streptomycin in clinical isolates of Mycobacterium tuberculosis by molecular methods.

Authors:  I Nachamkin; C Kang; M P Weinstein
Journal:  Clin Infect Dis       Date:  1997-05       Impact factor: 9.079

4.  Corneal blindness in a southern Indian population: need for health promotion strategies.

Authors:  R Dandona; L Dandona
Journal:  Br J Ophthalmol       Date:  2003-02       Impact factor: 4.638

5.  Utility of real-time PCR analysis for appropriate diagnosis for keratitis.

Authors:  Tomoyuki Inoue; Yuichi Ohashi
Journal:  Cornea       Date:  2013-11       Impact factor: 2.651

6.  Evaluation of corneal scraping smear examination methods in the diagnosis of bacterial and fungal keratitis: a survey of eight years of laboratory experience.

Authors:  Savitri Sharma; Derek Y Kunimoto; Usha Gopinathan; Sreedharan Athmanathan; Prashant Garg; Gullapalli N Rao
Journal:  Cornea       Date:  2002-10       Impact factor: 2.651

7.  Amplification of bacterial DNA using highly conserved sequences: automated analysis and potential for molecular triage of sepsis.

Authors:  K M McCabe; G Khan; Y H Zhang; E O Mason; E R McCabe
Journal:  Pediatrics       Date:  1995-02       Impact factor: 7.124

8.  Suppurative corneal ulceration in Bangladesh. A study of 142 cases examining the microbiological diagnosis, clinical and epidemiological features of bacterial and fungal keratitis.

Authors:  A A Dunlop; E D Wright; S A Howlader; I Nazrul; R Husain; K McClellan; F A Billson
Journal:  Aust N Z J Ophthalmol       Date:  1994-05

9.  Inhibition of PCR by aqueous and vitreous fluids.

Authors:  D L Wiedbrauk; J C Werner; A M Drevon
Journal:  J Clin Microbiol       Date:  1995-10       Impact factor: 5.948

10.  16S ribosomal DNA typing for identification of pathogens in patients with bacterial keratitis.

Authors:  C M Knox; V Cevellos; D Dean
Journal:  J Clin Microbiol       Date:  1998-12       Impact factor: 5.948

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  6 in total

1.  Effectiveness of 16S ribosomal DNA real-time PCR and sequencing for diagnosing bacterial keratitis.

Authors:  Daisuke Shimizu; Dai Miyazaki; Fumie Ehara; Yumiko Shimizu; Ryu Uotani; Koudai Inata; Shin-Ichi Sasaki; Yoshitsugu Inoue
Journal:  Graefes Arch Clin Exp Ophthalmol       Date:  2019-08-13       Impact factor: 3.117

2.  Comparison of culture, confocal microscopy and PCR in routine hospital use for microbial keratitis diagnosis.

Authors:  Jeremy J Hoffman; John K G Dart; Surjo K De; Nicole Carnt; Georgia Cleary; Scott Hau
Journal:  Eye (Lond)       Date:  2021-11-05       Impact factor: 4.456

3.  Utility of investigation for suspected microbial keratitis: a diagnostic accuracy study.

Authors:  Stephen Tuft; Catey Bunce; Surjo De; John Thomas
Journal:  Eye (Lond)       Date:  2022-01-31       Impact factor: 4.456

4.  16S Ribosomal RNA PCR Versus Conventional Diagnostic Culture in the Investigation of Suspected Bacterial Keratitis.

Authors:  Tobi F Somerville; Caroline E Corless; Henri Sueke; Timothy Neal; Stephen B Kaye
Journal:  Transl Vis Sci Technol       Date:  2020-12-01       Impact factor: 3.283

Review 5.  Diagnostic armamentarium of infectious keratitis: A comprehensive review.

Authors:  Darren S J Ting; Bhavesh P Gopal; Rashmi Deshmukh; Gerami D Seitzman; Dalia G Said; Harminder S Dua
Journal:  Ocul Surf       Date:  2021-11-13       Impact factor: 5.033

Review 6.  Diagnostic methods for the etiological assessment of infectious corneal pathology (Review).

Authors:  Mihail Zemba; Otilia-Maria Dumitrescu; Andreea-Elena Dimirache; Daniel Constantin Branisteanu; Florian Balta; Marian Burcea; Andreea Dana Moraru; Sinziana Gradinaru
Journal:  Exp Ther Med       Date:  2021-12-13       Impact factor: 2.447

  6 in total

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