Polymerase Chain Reaction and Its Application in the Diagnosis of Infectious Keratitis
Medical hypothesis discovery and innovation in ophthalmology,
Vol. 8 No. 3 (2019),
20 September 2019
,
Page 152-155
Abstract
PCR involves a repeating cycle of replication to amplify small segments of deoxyribonucleic acid (DNA). A novel application of this technique is microbial identification in infectious keratitis, one of the leading causes of blindness in the world. PCR is more sensitive than biological stains and culture, which are considered the current gold standards for diagnosing infectious keratitis. The diagnosis and treatment of infectious keratitis cost the United States millions of dollars in health expenditure. PCR may help offset that cost by allowing for individualized disease management and screening for multiple antibiotic-resistant genes. While beneficial, PCR demonstrates lower specificity rates compared to culture and stain, indicating its shortcomings; this can be overcome by performing PCR after narrowing the pool of potential microorganisms. This article examines the clinical utility of PCR in cases of infectious keratitis by evaluating its reliability, validity, associated costs, and indications.
References
Collier SA, Gronostaj MP, MacGurn AK, Cope JR, Awsumb KL, Yoder JS, et al. Estimated burden of keratitis--United States, 2010. MMWR Morb Mortal Wkly Rep. 2014;63:1027-30. pmid: 25393221
Austin A, Lietman T, Rose-Nussbaumer J. Update on the Management of Infectious Keratitis. Ophthalmology. 2017;124:1678-89. doi: 10.1016/j.ophtha.2017.05.012 pmid: 28942073
Pascolini D, Mariotti SP. Global estimates of visual impairment: 2010. Br J Ophthalmol. 2012;96:614-8. doi: 10.1136/bjophthalmol-2011-300539 pmid: 22133988
Ung L, Bispo PJM, Shanbhag SS, Gilmore MS, Chodosh J. The persistent dilemma of microbial keratitis: Global burden, diagnosis, and antimicrobial resistance. Surv Ophthalmol. 2019;64:255-71. doi: 10.1016/j.survophthal.2018.12.003 pmid: 30590103
Whitcher JP, Srinivasan M. Corneal ulceration in the developing world--a silent epidemic. Br J Ophthalmol. 1997;81:622-3. doi: 10.1136/bjo.81.8.622 pmid: 9349145
Kim E, Chidambaram JD, Srinivasan M, Lalitha P, Wee D, Lietman TM, et al. Prospective comparison of microbial culture and polymerase chain reaction in the diagnosis of corneal ulcer. Am J Ophthalmol. 2008;146:714-23, 23 e1. doi: 10.1016/j.ajo.2008.06.009 pmid: 18707670
Eleinen KG, Mohalhal AA, Elmekawy HE, Abdulbaki AM, Sherif AM, El-Sherif RH, et al. Polymerase chain reaction-guided diagnosis of infective keratitis - a hospital-based study. Curr Eye Res. 2012;37:1005-11. doi: 10.3109/02713683.2012.698357 pmid: 22746322
Hematian A, Sadeghifard N, Mohebi R, Taherikalani M, Nasrolahi A, Amraei M, et al. Traditional and Modern Cell Culture in Virus Diagnosis. Osong Public Health Res Perspect. 2016;7:77-82. doi: 10.1016/j.phrp.2015.11.011 pmid: 27169004
Waters DL, Shapter FM. The polymerase chain reaction (PCR): general methods. Methods Mol Biol. 2014;1099:65-75. doi: 10.1007/978-1-62703-715-0_7 pmid: 24243196
Ishmael FT, Stellato C. Principles and applications of polymerase chain reaction: basic science for the practicing physician. Annals of Allergy, Asthma & Immunology. 2008;101:437-43. doi: 10.1016/s1081-1206(10)60323-7
Maharana PK, Sharma N, Nagpal R, Jhanji V, Das S, Vajpayee RB. Recent advances in diagnosis and management of Mycotic Keratitis. Indian J Ophthalmol. 2016;64:346-57. doi: 10.4103/0301-4738.185592 pmid: 27380973
Markoulatos P, Siafakas N, Moncany M. Multiplex polymerase chain reaction: a practical approach. J Clin Lab Anal. 2002;16:47-51. doi: 10.1002/jcla.2058 pmid: 11835531
Strommenger B, Kettlitz C, Werner G, Witte W. Multiplex PCR assay for simultaneous detection of nine clinically relevant antibiotic resistance genes in Staphylococcus aureus. J Clin Microbiol. 2003;41:4089-94. doi: 10.1128/jcm.41.9.4089-4094.2003 pmid: 12958230
Chambers HF. Methicillin resistance in staphylococci: molecular and biochemical basis and clinical implications. Clin Microbiol Rev. 1997;10:781-91. pmid: 9336672
Zhao G, Zhai H, Yuan Q, Sun S, Liu T, Xie L. Rapid and sensitive diagnosis of fungal keratitis with direct PCR without template DNA extraction. Clin Microbiol Infect. 2014;20:O776-82. doi: 10.1111/1469-0691.12571 pmid: 24471925
Goh JWY, Harrison R, Hau S, Alexander CL, Tole DM, Avadhanam VS. Comparison of In Vivo Confocal Microscopy, PCR and Culture of Corneal Scrapes in the Diagnosis of Acanthamoeba Keratitis. Cornea. 2018;37:480-5. doi: 10.1097/ICO.0000000000001497 pmid: 29256983
Walker B, Powers-Fletcher MV, Schmidt RL, Hanson KE. Cost-Effectiveness Analysis of Multiplex PCR with Magnetic Resonance Detection versus Empiric or Blood Culture-Directed Therapy for Management of Suspected Candidemia. J Clin Microbiol. 2016;54:718-26. doi: 10.1128/JCM.02971-15 pmid: 26739159
Gopinathan U, Sharma S, Garg P, Rao GN. Review of epidemiological features, microbiological diagnosis and treatment outcome of microbial keratitis: experience of over a decade. Indian J Ophthalmol. 2009;57:273-9. doi: 10.4103/0301-4738.53051 pmid: 19574694
Witt N, Rodger G, Vandesompele J, Benes V, Zumla A, Rook GA, et al. An assessment of air as a source of DNA contamination encountered when performing PCR. J Biomol Tech. 2009;20:236-40. pmid: 19949694
Ting DSJ, Settle C, Morgan SJ, Baylis O, Ghosh S. A 10-year analysis of microbiological profiles of microbial keratitis: the North East England Study. Eye (Lond). 2018;32:1416-7. doi: 10.1038/s41433-018-0085-4 pmid: 29610521
Stevenson G, Vemulapalli R, Wu C. New PCR Tests at ADDL 2002 [June 6, 2019]. Available from: https://www.addl.purdue.edu/newsletters/2002/fall/pcr.shtml.
Beal SG, Tremblay EE, Toffel S, Velez L, Rand KH. A Gastrointestinal PCR Panel Improves Clinical Management and Lowers Health Care Costs. J Clin Microbiol. 2018;56. doi: 10.1128/JCM.01457-17 pmid: 29093106
Keay L, Edwards K, Naduvilath T, Taylor HR, Snibson GR, Forde K, et al. Microbial keratitis predisposing factors and morbidity. Ophthalmology. 2006;113:109-16. doi: 10.1016/j.ophtha.2005.08.013 pmid: 16360210
Van Cleeff M, Kivihya-Ndugga L, Githui W, Ng'ang'a L, Kibuga D, Odhiambo J, et al. Cost-effectiveness of polymerase chain reaction versus Ziehl-Neelsen smear microscopy for diagnosis of tuberculosis in Kenya. The International Journal of Tuberculosis and Lung Disease. 2005;9:877-83.
Scherer LC, Sperhacke RD, Ruffino-Netto A, Rossetti ML, Vater C, Klatser P, et al. Cost-effectiveness analysis of PCR for the rapid diagnosis of pulmonary tuberculosis. BMC Infect Dis. 2009;9:216. doi: 10.1186/1471-2334-9-216 pmid: 20043842
Inoue T, Kawashima R, Suzuki T, Ohashi Y. Real-time polymerase chain reaction for diagnosing acyclovir-resistant herpetic keratitis based on changes in viral DNA copy number before and after treatment. Arch Ophthalmol. 2012;130:1462-4. doi: 10.1001/archophthalmol.2012.1176 pmid: 23143448
Jung HS, Tsongalis GJ, Lefferts JA. Development of HLA-B*57:01 Genotyping Real-Time PCR with Optimized Hydrolysis Probe Design. J Mol Diagn. 2017;19:742-54. doi: 10.1016/j.jmoldx.2017.05.002 pmid: 28732216
Houssin T, Cramer J, Grojsman R, Bellahsene L, Colas G, Moulet H, et al. Ultrafast, sensitive and large-volume on-chip real-time PCR for the molecular diagnosis of bacterial and viral infections. Lab Chip. 2016;16:1401-11. doi: 10.1039/c5lc01459j pmid: 26952334
Dworkin LL, Gibler TM, Van Gelder RN. Real-time quantitative polymerase chain reaction diagnosis of infectious posterior uveitis. Arch Ophthalmol. 2002;120:1534-9. doi: 10.1001/archopht.120.11.1534 pmid: 12427068
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