Ilamathy, M (2017) A Study on Performance Evaluation and Utility of RT-LAMP (Reverse transcription loop-mediated isothermal amplification assay) for early detection of Dengue fever and Molecular characterization of Dengue virus by PCR, in a tertiary care hospital. Masters thesis, Madras Medical College, Chennai.
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Abstract
INTRODUCTION: Dengue virus is a mosquito-borne flavivirus and the most widely prevalent arbovirus in tropical and subtropical region. There are four distinct serotypes, DEN-1, DEN-2, DEN-3, and DEN-4, which produce a spectrum of illness ranging from inapparent infection to moderate febrile illness and severe and fatal hemorrhagic disease. Early, sensitive and specific diagnosis is paramount for etiologic investigation, patient management, prevention of complications and disease control in the community OBJECTIVES: 1. The performance of RT-LAMP with regard to sensitivity, specificity and cost would be compared with RT-PCR. 2. Usefulness of RT-LAMP in early diagnosis of Dengue would be evaluated in comparison with serological assays. 3. To characterize and to study on most prevalent serotype of Dengue virus in our centre. MATERIALS AND METHODS: A total of 100 acute phase serum samples from outpatients and inpatients with a clinical diagnosis of Dengue fever attending RGGGH, Chennai, Tamilnadu were included in the study. All the samples were tested for the presence of DENV specific RNA by RT-PCR, RT- LAMP and NS1 antigen by ELISA. IgM ELISA was done in serum samples from the study group patients, in whom a second samples could be obtained (82 patients) after one week of fever. RESULTS: In a total of 100 samples,RT-LAMP was positive in 44% and RT-PCR was positive 36%of the samples. All of the RT-PCR positive samples were positive by RT-LAMP.In addition RT-LAMP showed positivity in 8 samples which were negative by RT-PCR. The RT-LAMP assay demonstrated higher sensitivity than conventional RT-PCR by picking up 8 additional positive cases. The higher sensitivity and specificity of the RT-LAMP reaction are due to continuous amplification under isothermal conditions and by employing six primers that specifically recognize eight distinct regions of the target. The most predominant serotype was DEN-2 (94%), DEN- 3 (6%). IgM ELISA was positive in 54% of the patients. CONCLUSION: RT-LAMP assay is a rapid, accurate and cost effective diagnostic tool for identification of Dengue virus. Due to its simplicity not requiring any sophisticated equipment it has been emerging as a potential alternative to conventional RT-PCR for clinical diagnosis and surveillance of DENV in our country.
| Item Type: | Thesis (Masters) |
|---|---|
| Additional Information: | Reg.No.201414004 |
| Uncontrolled Keywords: | Dengue fever, Reverse Transcription Loop Mediated Isothermal Amplification Assay (RT-LAMP), Reverse Transcription Polymerase Chain Reaction (RT-PCR). |
| Subjects: | MEDICAL > Microbiology |
| Depositing User: | Subramani R |
| Date Deposited: | 18 Jul 2020 17:23 |
| Last Modified: | 19 Jul 2020 04:15 |
| URI: | http://repository-tnmgrmu.ac.in/id/eprint/12545 |
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