Sensitive and Rapid Polymerase Chain Reaction Based Diagnosis of Mycotic Keratitis Through Single Stranded Conformation Polymorphism

Kumar, Manish; Mishra, Niraj; Shukla, Praveen Kumar

doi:10.1016/j.ajo.2005.05.019

Cited by 31 publications

(24 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…21 These unusual fungi identified by PCR might be uncultured by routine media, as has been reported from previous studies. 20,22,23 A. alternata has been reported as a corneal pathogen by PCR. 24 In this series, A. alternata was identified in one case by PCR analysis whereas Fusarium species was grown on a routine culture.…”

Section: Discussionmentioning

confidence: 99%

Prospective comparison between conventional microbial work-up vs PCR in the diagnosis of fungal keratitis

Tananuvat

Salakthuantee

Vanittanakom

et al. 2012

Eye

View full text Add to dashboard Cite

Purpose The PCR was compared with routine microbial studies for the detection of fungal pathogens in clinically suspected fungal keratitis. Methods A prospective nonrandomized study was undertaken at a tertiary eye care centre to evaluate 30 eyes of 30 patients with presumed fungal keratitis, both fresh and treated. Corneal scrapings were performed on each patient. The specimens were analysed by a semi-nested PCR assay using fungalspecific primers. PCR products were cloned and sequenced for identification, and compared with a conventional microbial work-up (smear and culture). Results Of the 30 samples, the PCR showed positivity in 93.3%, culture in 40%, and potassium hydroxide in 20%. Of the 28 PCR-positive cases, 12 were culture-positive and 16 were culture-negative. Two samples were both PCR and culture test negative. Culture-negative samples were PCR-positive in 16 of 18 (88.9%) cases. The PCR did not yield any false-negative findings in a culturepositive specimen. Both common and uncommon aetiologic fungi have been identified by DNA sequencing analysis. Conclusion The PCR was able to detect fungal DNA in a high proportion of culturenegative cases. Technical considerations of the PCR process include extraction of artifacts and amplification of non-pathogenic DNA. Nonetheless, our findings suggest that the PCR can be a useful adjunct to smear and culture in the rapid diagnosis of fungal keratitis, particularly in cases of failed detection from routine procedures.

show abstract

Section: Discussionmentioning

confidence: 99%

Prospective comparison between conventional microbial work-up vs PCR in the diagnosis of fungal keratitis

Tananuvat

Salakthuantee

Vanittanakom

et al. 2012

Eye

View full text Add to dashboard Cite

show abstract

“…The ITS sequence of the type strain of B. hawaiiensis showed similarities of 93.7 and 94.8% with those of the type strains of B. australiensis and B. spicifera, respectively. A BLAST search comparing ITS sequences is the procedure usually used in recent years to confirm Bipolaris species identification (3,6,9,16,18). However, this procedure may not be useful when comparisons are made with inaccurate sequences or when sequences of authentic strains of the species to which the problem isolate belongs have not been deposited.…”

Section: Discussionmentioning

confidence: 99%

Diversity of Bipolaris Species in Clinical Samples in the United States and Their Antifungal Susceptibility Profiles

et al. 2012

View full text Add to dashboard Cite

“…Kumar et al [21,22] have investigated the use of PCR in detecting fungal pathogens in keratitis, and samples from 4 patients with mycotic keratitis were examined, along with other samples obtained directly from fungal cultures. They showed that the PCR combined with single-stranded conformational analysis allow for rapid and precise identification of unusual mycotic pathogens.…”

Section: Discussionmentioning

confidence: 99%

Comparative Study of Gram Stain, Potassium Hydroxide Smear, Culture and Nested PCR in the Diagnosis of Fungal Keratitis

Badiee¹,

Nejabat

Alborzi³

et al. 2010

Ophthalmic Res

View full text Add to dashboard Cite

Objective: This study seeks to evaluate the efficacy and practicality of the molecular method, compared to the standard microbiological techniques for diagnosing fungal keratitis (FK). Methods: Patients with eye findings suspected of FK were enrolled for cornea sampling. Scrapings from the affected areas of the infected corneas were obtained and were divided into two parts: one for smears and cultures, and the other for nested PCR analysis. Results: Of the 38 eyes, 28 were judged to have fungal infections based on clinical and positive findings in the culture, smear and responses to antifungal treatment. Potassium hydroxide, Gram staining, culture and nested PCR results (either positive or negative) matched in 76.3, 42.1, 68.4 and 81.6%, respectively. Conclusion: PCR is a sensitive method but due to the lack of sophisticated facilities in routine laboratory procedures, it can serve only complementarily and cannot replace conventional methods.

show abstract

Sensitive and Rapid Polymerase Chain Reaction Based Diagnosis of Mycotic Keratitis Through Single Stranded Conformation Polymorphism

Cited by 31 publications

References 25 publications

Prospective comparison between conventional microbial work-up vs PCR in the diagnosis of fungal keratitis

Prospective comparison between conventional microbial work-up vs PCR in the diagnosis of fungal keratitis

Diversity of Bipolaris Species in Clinical Samples in the United States and Their Antifungal Susceptibility Profiles

Comparative Study of Gram Stain, Potassium Hydroxide Smear, Culture and Nested PCR in the Diagnosis of Fungal Keratitis

Contact Info

Product

Resources

About