1.Molecular identification of Leishmania tropica infections in patients with cutaneous leishmaniasis from an endemic central of Iran
Gilda Eslami ; Bahador Hajimohammadi ; Abbas Ali Jafari ; Farzaneh Mirzaei ; Mostafa Gholamrezai ; Hossein Anvari ; Ali Gilda Eslami ; Bahador Hajimohammadi ; Abbas Ali Jafari ; Farzaneh Mirzaei ; Mostafa Gholamrezai ; Hossein Anvari ; Ali Khamesipour
Tropical Biomedicine 2014;31(4):592-599
The most common form of the disease is cutaneous leishmaniasis (CL) which is a
public health and social problem in many countries especially Iran. In endemic areas where
other diseases with similar clinical symptoms occur, definitive diagnosis of CL is very important.
The detection and identification of Leishmania in infected patients is crucial for achieving a
correct treatment and prognosis. To our knowledge, this is the first comprehensive study in
terms of geographical distribution and molecular identification of Leishmania tropica isolates
in central of Iran. This study was performed between 2010 and 2011, during which 218 CL
suspected patients referred to Shahid Sadoughi University of Medical Sciences in Yazd, Iran
for confirmation were examined. After microscopic analysis, DNA extraction was performed
for identification. The molecular target region was ITS1 gene. Results showed that out of 218
isolates, 102 (46.8%) samples were positive for Leishman body using molecular assay. After
PCR-RFLP, analysis identified 50 (49.01%) samples as L. major and 52 (50.98%) as L. tropica.
Two samples showed a different pattern that were reported as unknown. Among L. tropica,
six different isolates were identified in this endemic area. Finally, this study showed
heterozygosity among L. tropica isolates in this endemic area such as some other studies
from the world. This heterozygosity among the strains may suggest a sexual recombination or
genetic exchange between strains.
2.Rapid detection of MDR-Mycobacterium tuberculosis using modified PCR-SSCP from clinical Specimens.
Imani Fooladi Abbas ALI ; Farzam BABAK ; Mousavi Seyed FAZLOLLAH ; Jonaidi Jafari NEMATOLLAH
Asian Pacific Journal of Tropical Biomedicine 2014;4(Suppl 1):S165-70
OBJECTIVETo design a rapid test to detect the rifampin (RIF) and isoniazid (INH) resistant mutant based on polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technique that analyzes the katG, rpoB genes.
METHODSBiochemical test as well as IS6110 targeting PCR revealed 103 clinical samples were tuberculosis. To determine the susceptibility of isolates to anti TB drugs, the proportional method was used. Mutations presented within the amplified products of the katG, rpoB genes were evaluated by SSCP.
RESULTSUsing proportional method, 12 (11.6%) and 9 (8.7%) isolates were resistant respectively to INH and RIF and 9 (8.7%) isolates showed resistance to both drug (multi-drug resistant tuberculosis). Three (2.9%) multi-drug resistant tuberculosis and two INH resistant isolates were detected by the PCR-SSCP and sequencing. The sensitivity and specificity of PCR-SSCP for multi-drug resistant isolates were 33% and 100%, respectively.
CONCLUSIONSComplete agreement between SSCP and sequencing can indicate that resistance-associated mutations have occurred in other genes except our considered genes.