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Fast Method for Diagnosis of Leishmania by PCR and FLASH PCR
Alireza Moradabadi, Alireza Farsinezhad, Maryam Fekri Soofiabadi,
Volume 19, Issue 11 (2-2017)
Abstract

Background: Leishmaniasis is a protozoan parasitic disease and a global health problem. The aim of this study is to diagnose the parasitic infection in humans for epidemiological identification and providing control programs using proprietary co-designed primers in three species of Leishmania.

Materials and Methods: 30 common Leishmania isolates were gathered from different centers in Iran. Having been cultured in RPMI-1640 Medium, DNA was extracted and the gene   ITS2-rRNA was amplified by PCR. The amplicons were examined by electrophoresis on agarose gel 2%. Also, in FLASH PCR method, a specific probe and florence colour were used to investigae the amplicon existence on sample.

Results: The results of the investigations by PCR and FLASH PCR methods show that these methods are sensitive and specific for diagnosis of Leishmania

Conclusion: In this study, identification of Leishmania parasite using specific primer pairs was successful and TaqMan could be one of the most sensitive diagnostic methods to identify parasite load for the ITS2 region of Leishmania.


Comparison of Detection of Clinical Isolates of Mycobacterium Tuberculosis by Flash PCR and Conventional Culture Method
Ali Reza Morad Abadi, Mohammad Arjomandzadegan, Navid Emami, Manijeh Kahbazi, Azam Ahmadi, Saeed Falahat, Seyyed Hossein Hosseini, Mehdi Kargaran, Parisa Khosravi,
Volume 21, Issue 4 (8-2018)
Abstract
Background and Aim: Ziehl Nelson staining, fluorescent and also culture are the standard methods for the diagnosis of tuberculosis. In this study, the performance of conventional cultivation methods was compared with Flash PCR.
Materials and Methods: A total of 56 sputum samples from patients with suspected tuberculosis in Tuberculosis Center of Arak city were collected and Ziehl–Neelsen and culture in Löwenstein–Jensen medium were accomplished. Moreover, DNA from all of the 56 sputum samples was extracted by Chelex100 method. Molecular evaluation was accomplished by Flash PCR kit containing probes and primers for gene amplification IS6110. Positive and negative controls together with samples were used in a MTC410 apparatus for amplification. FD-12 apparatus was used to evaluate the results. In addition, electrophoresis on agarose was used for confirmation of the results.
Findings: From 56 sputum samples of suspected TB patients, 20 samples were positive and 36 samples were negative on microscopic evaluation and culture methods. FLASH-PCR molecular analysis showed that all of 20 positive samples were positive in molecular methods, too. On the other hand, three of sputum samples that were negative by culture and staining were positive in FLASH-PCR method. One of these 3 patients, received Isoniazid, pyrazinamide and ethambutol antibiotic by responsible medicine. All results were confirmed using conventional electrophoresis.
Conclusion: In some negative samples, possibly because of the small number of bacteria in sample or a defect in the sampling, the Flash PCR may due good advantages. Therefore, due to the low cost, this method is recommended for routine use.

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