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[Rapid pathogens diagnosis of infected keratitis and endophthalmitis using two steps polymerase chain reaction]
1Department of Ophthalmology, the Affiliated Hospital of Nantong Medical College, Nantong 226001, China. Guanhj@public.nt.js.cn
Objective:
To establish a rapid diagnosis technique of the pathogens of infected keratitis and endophthalmitis.
Methods:
Fifty-one corneal scraping, tear and 16 vitreous specimens were obtained from 67 cases of infected keratitis and endophthalmitis. Multiplex polymerase chain reaction (MPCR) was performed to differentiate the genes of bacteria, fungi and HSV using their own universal primers. Special-primers PCR were carried out for MPCR positive specimens to detect the genes of common bacteria and fungi respectively. The results were compared with microbe culture and clinical diagnosis.
Results:
In 67 clinical samples, 52 (77.6%) were positive at least one kind of pathogen by MPCR (two positive pathogens were 8 cases). SPCR revealed that 7 bacteria (25.0%) out of 28 bacteria positive specimens were Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa. Eight fungi (53.3%)of 15 fungi positive specimens was Candida, Aspergillus, Fusarium. The culture positive rates were 31.3% (21 cases), among them 1 case was found positive for two pathogens. MPCR positive rate was statistically higher than the rate that in culture (chi(2) = 13.50, P < 0.005). The kinds of bacteria and fungi of SPCR positive were as the same as bacteria and fungi of culture positive. Consistency of clinical diagnosis and experimental evidences of infected keratitis was 54.8%. The time for microbe culture needs 72 to 96 hours, but MPCR and SPCR needs 8 hours.
Conclusion:
Two steps PCR is rapid, sensitive method to detect the pathogens of infected keratitis and endophthalmitis.
Insights
Multiplex polymerase chain reaction (MPCR) and special-primers PCR offer a rapid and sensitive method for diagnosing infected keratitis and endophthalmitis. This technique significantly outperforms traditional microbe culture in pathogen detection speed and accuracy.
Area of Science:
- Ophthalmology
- Microbiology
- Molecular Biology
Context:
- Infectious keratitis and endophthalmitis are serious ocular conditions requiring prompt diagnosis and treatment.
- Current diagnostic methods, such as microbe culture, can be slow, delaying critical therapeutic interventions.
- Rapid and accurate pathogen identification is crucial for effective management and visual outcome preservation.
Purpose:
- To establish and evaluate a rapid diagnostic technique for identifying pathogens responsible for infectious keratitis and endophthalmitis.
- To compare the sensitivity and speed of multiplex polymerase chain reaction (MPCR) and special-primers PCR (SPCR) against conventional microbe culture and clinical diagnosis.
Summary:
- MPCR and SPCR were performed on 67 clinical specimens (corneal scrapings, tears, vitreous).
- The PCR-based methods detected pathogens in 77.6% of samples, significantly higher than microbe culture (31.3%).
- MPCR and SPCR identified specific bacteria and fungi, demonstrating high concordance with culture results, and provided results within 8 hours compared to 72-96 hours for culture.
Impact:
- This two-step PCR approach offers a significantly faster and more sensitive diagnostic tool for ocular infections.
- The enhanced diagnostic capability can lead to earlier treatment initiation, potentially improving patient outcomes and reducing vision loss.
- The study highlights the clinical utility of molecular techniques in managing complex eye infections.
