Detection of Leishmania major using PCR-ELISA

Authors

  • Afieh Samimi Department of Biotechnology, Faculty of New Technologies, Golestan University of Medical Sciences, Iran
  • Beniamin Talebi Private veterinary physician, Gorgan,Iran
  • Faramarz Koohsar Laboratory Sciences Research Center, Faculty of Paramedicine, Golestan University of Medical Sciences, Gorgan, Iran
  • Fatemeh Mesgarian Department of Medical Parasitology, Gonbad-e Kavus Health Center, Golestan University of Medical Sciences, Gorgan, Iran
  • khodaberdi kalavi Laboratory Sciences Research Center, Faculty of Paramedicine, Golestan University of Medical Sciences, Gorgan, Iran
  • Oghol Niaz Jorjani Laboratory Science Research Center, Faculty of Paramedicine, Golestan University of Medical Sciences, Gorgan, Iran
  • Zohreh Sharifi Professor, Department of Virology, Iranian Blood Transfusion Organization, Tehran, Iran
Abstract:

Background and objectives: Cutaneous leishmaniasis is endemic in most areas of Iran, and the diagnosis of its species is essential for controlling the disease. Leishmania major is the causative agent for cutaneous leishmaniasis in humans. Molecular methods are generally more sensitive than microscopic methods. The present study aimed to use a polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA) technique for detecting live L. major from wounds of patients with cutaneous leishmaniasis. Methods: In the present study, a standard strain of L. major promastigotes was used as the positive control for purification of DNA. The Novy–MacNeal–Nicolle and RPMI-1640 media were used for reproduction of parasites. DNA was isolated from specimens taken from 35 patients with suspected cutaneous leishmaniasis whose disease was confirmed by direct smear method. The PCR-ELISA technique was later applied by using the standard strain, patient specimens, and primers specific for the 18s rRNA. Results: Out of 35 patients, 17 (48.6%) were male and 18 (51.4%) were female. In addition, 8.6% of the patients lived in the Gonbad-e Kavus County, while all patients had been infected in villages around Gonbad-e Kavus. Of 35 patients with confirmed cutaneous leishmaniasis according to the direct smear method, 31 patients (86.31%) had leishmaniasis based on the PCR method and the PCR-ELISA methods. Conclusion: Based on the results, the PCR-ELISA method is more sensitive and accurate for detecting L. major.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

Diagnosis of infections with Leishmania infantum using PCR-ELISA.

On the basis of partial amplification of a cloned fragment of kDNA of Leishmania infantum which is specific for this species, we developed a PCR-ELISA technique which avoids the problems associated with classical diagnostic techniques. This technique was tested on 33 L. infantum strains from 19 different zymodemes, which were recognized equally. It was also used on human and canine clinical sam...

full text

Optimization of PCR-ELISA in Detection of Human Cytomegalovirus Infection

Abstract Background and Objective: Human Cytomegalovirus (CMV) is an important cause of congenital viral infection that can lead to serious diseases and complications in infants. Application of rapid, sensitive, and specific HCMV detection methods is necessary for congenital infection detection. We aimed to optimize the use of PCR and ELISA for detection of HCMV in infants. Material and Methods...

full text

detection of leishmania major in naturally infected sand flies using semi nested-pcr

background: the aim of this study was to assess leishmania infection in sand fly species from areas where leishmaniasis is endemic. this is important for prediction of the risk and expansion of the disease. methods: in this cross-sectional study we used a pcr-based method for detection of leishmania minicircle dna within individual sand flies from orzoieh, a new endemic leishmaniasis focus in s...

full text

Validation of a β-ME ELISA for Detection of Anti Leishmania donovani Antibodies in Eastern Sudan

Background: A β-mercaptoethnol (β-ME)-treated promastigote antigen of L. donovani was successfully employed in direct agglutination test (DAT) for the diagnosis of visceral leishmaniasis (VL). Objective: The β-ME-treated antigen was further incorporated into an enzyme-linked immunosorbent assay set-up (β-ME ELISA) and evaluated for VL diagnosis against outcome of reference freeze-dried DAT (FD-...

full text

Comparison between conventional PCR and PCR - ELISA for detection of Brucella melitensis

Molecular detection techniques are believed to be key tools for both prevention and treatment follow up of brucellosis within live stock and human beings. Consequently rapid, reliable, easy to perform and automated systems for Brucella detection are urgently needed to allow early diagnosis and adequate antibiotic therapy in time. Brucellosis is a worldwide re-emerging zoonosis causing high econ...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 16  issue 3

pages  24- 29

publication date 2022-05

By following a journal you will be notified via email when a new issue of this journal is published.

Keywords

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023