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Optimization of Loop-Mediated Isothermal Amplification (LAMP) method for detection of herpes simplex virus type 1 and 2
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Asmaa Bahrami1   , Mohammad Hasan Shahhosseiny 2, Kayhan Azadmanesh3 , Zahra Noormohamadi4 , Elham Moslemi5 , Farzaneh Jadali6 |
1- Islamic Azad University, Science & Research Branch, Tehran
2- Islamic Azad University, Shahr-e- Qods Branch , shahhosseiny@yahoo.com
3- Pasteur Institute of Iran
4- Islamic Azad University, Science & Research Branch
5- Islamic Azad University, East Tehran Branch
6- Pathology Dept., Mofid Hospital of Tehran |
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Abstract: (15221 Views) |
| ABSTRACT
Background and Aim: Type 1 and type 2 herpes simplex (HSV) virus cause infection of central nervous system (encephalitis) in human. The molecular techniques are the best methods for detection of HSV. In this study we evaluated the novel molecular technique of LAMP for detection of HSV-1 and HSV-2.
Material and Methods: In this experimental study 184 cerebrospinal fluid (CSF) samples were collected from Mofid Hospital. DNA of every sample was extracted by use of Sinagen DNP Kit. Based on the HSV DNA polymerase gene, a set of 6 primers were designed and sensitivity and specificity of this method were determined. By adding SYBER Green, LAMP product was identified. The results of LAMP method were compared to those of PCR by chi-square test.
Results: Sensitivity of LAMP method determined to be 5 copies/ tube and sensitivity of PCR method determined to be 50 copies/ tube. Both LAMP and PCR methods showed 100% specificity for detection of HSV type 1 and type2. Among 184 samples, 60 samples were positive by LAMP but 45 samples were positive by PCR method. Sensitivity of LAMP was 10 times higher than that of PCR. Comparison of the results of the two methods by means of chi-square test showed a significant difference (p‹0.05).
Conclusion: LAMP method had high sensitivity and specificity for detection of type 1 and type 2 HSV in CSF samples.
Key word: HSV, LAMP, CSF
Conflict of Interest: Nill
Received: Nov 22, 2010 Accepted: Jan 14, 2011 |
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| Keywords: HSV, LAMP, CSF |
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Full-Text [PDF 1671 kb]
(3826 Downloads)
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Type of Study: Original Research |
Subject:
General
Received: 2011/05/10 | Published: 2011/05/15
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