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qPCR Is a Sensitive and Rapid Method for Detection of Cytomegaloviral DNA in Formalin-fixed, Paraffin-embedded Biopsy Tissue
Published on: July 9, 2014
Cytomegalovirus infection in patients attending a tertiary care hospital - single center experience
Bashir Ahmad Fomda1, Munaza Aman1, Sanam Wani1
1Department of Microbiology, Sher-I-Kashmir Institute of Medical Sciences, Soura, Srinagar.
Insights
Human cytomegalovirus (CMV) disease is a significant public health concern in India. This study found CMV infection in 21% of suspected cases, highlighting the need for early detection using PCR.
Area of Science:
- Virology
- Infectious Diseases
- Molecular Diagnostics
Background:
- Human cytomegalovirus (CMV) infection is a critical public health issue, particularly under-addressed in India.
- Previous research focused on CMV seroprevalence, necessitating studies on the actual disease burden.
Purpose of the Study:
- To determine the true prevalence and magnitude of human cytomegalovirus (CMV) disease in a patient cohort.
- To evaluate the utility of real-time PCR for diagnosing CMV disease.
Main Methods:
- Real-time PCR was employed to detect human cytomegalovirus (CMV) DNA in 181 patient samples.
- Simultaneous detection of CMV DNA and an internal control used specific probes (FAM™ and JOE™).
- Positive results were defined by amplification curves crossing a threshold within 36 cycles.
Main Results:
- Human cytomegalovirus (CMV) infection was detected in 21% of the analyzed samples.
- The most frequent clinical presentation was meningitis/meningoencephalitis.
- Infections were most prevalent in infants (18.2%) and post-renal transplant patients (2.7%), with detection in urine (17.1%) and serum (3.8%).
Conclusions:
- Real-time PCR plays a crucial role in the early diagnosis of human cytomegalovirus (CMV) disease.
- Timely diagnosis is essential to prevent irreversible complications associated with delayed treatment.
Background And Objectives:
Human cytomegalovirus infection poses an important public health issue. This issue in India has not received enough attention. The majority of research workers have highlighted the seroprevalence of human cytomegalovirus. Hence this study was conducted to find out true magnitude of human cytomegalovirus disease.
Materials And Methods:
Samples from 181 patients with suspected human cytomegalovirus disease were analyzed for human cytomegalovirus. DNA extraction was followed by real-time PCR. Human cytomegalovirus DNA-specific probes, fluorophore FAM™ and fluorophore JOE™ were utilized to detect human cytomegalovirus specific DNA and internal control at the same time. After completion of the assay, fluorescent growth curves were examined, and the response growth curves passing the threshold line in less than 36 cycles were deemed to be positive. All relevant clinical, demographic, and epidemiological information of the patients was also recorded.
Results:
The most common clinical presentation was meningitis/meningoencephalitis. Out of the total samples, human cytomegalovirus infection was detected in 21% of the samples. Most positive samples were from infants (18.2%), followed by post-renal transplant cases (2.7%). Human cytomegalovirus was detected in urine samples (17.1%) followed by serum (3.8%). Four out of the 14 CSF samples were tested for other viruses as well, and they were positive for EBV (n=1, 7%), enterovirus (n=2, 14%), and varicella zoster virus (n=1, 7%).
Conclusion:
PCR has a significant role in the detection of human cytomegalovirus disease at an early stage to avoid irreversible sequelae of late diagnosis.
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