Showing 2 results for Pcr-Elisa
M Talkhabifard, M, N Javid, N, A Moradi, A, A Ghaemi, A, A Tabarraei, A,
Volume 8, Issue 5 (1-2015)
Abstract
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: PCR–ELISA was performed by using specific primers and probe for detection of the HCMV glycoprotein B gene. First, the extracted DNA from urine samples and controls were labeled by digoxigenin during DIG-labeling PCR. After that, Biotin-labeled probe captured the DIG-labeled PCR products. The probe-PCR product hybrid is immobilized on a streptavidin-coated Microtiter plate, and detection was confirmed by proxidase-conjugated anti-digoxigenin antibody, and calorimetric substrate. Results: The clinical Human CMV strains isolated from16 patients were detected by this method. The optimized PCR-ELISA method was able to detect less than100 copies of HCMV genome. There was no non-specific reaction. Conclusion: PCR-ELISA can be applied as a sensitive, specific and reliable method for Semi-quantitative CMV detection in clinical samples. Keywords: Cytomegalovirus, Glycoprotein B, PCR-ELISA, Semi-Quantitative
Afieh Samimi, Oghol Niaz Jorjani, Zohreh Sharifi, Faramarz Koohsar, Khodaberdi Kalavi, Fatemeh Mesgarian, Beniamin Talebi ,
Volume 16, Issue 3 (5-2022)
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.
