رسائل ماجيستير
English
2021
Polymorphism of Toll-Like Receptors Genes of Susceptibility Patients with Pulmonary Aspergillosis
فرح محمد سعيد صادق الجبوري, كوثر محمد علي حسن
كلية العلوم للبنات-جامعة بابل · العراق
This study aimed to determine the occurrence of Aspergillus species among patients with lung problems, by means of phenotypic examination and molecular identification, as well as detection for some polymorphisms of Toll-like receptor genes of patients.
This study was carried out during the period from October 2020 to February 2021. A total of 128 samples of sputum and bronchial lavage fluid for both sexes and different age groups ranged between (15–83) were collected from patients suffering from lung infection, and they were complaining of symptoms indicative of lung disease who were admitted to the Specialized Center for Chest Diseases and Taiba Hospital in Al Hillah city.
As 102 samples were sputum and 26 samples of lung lavage fluid, and after the collection, each sample was cultured on the (SDA) and (PDA) and examined microscopically and grossly.
The present study showed that the percentage of samples that were positive culture in BAL was 24 (18.7%), while positive culture in sputum showed 93 (72.6%), and the total number of fungal colonies isolated was 445, while the total number of yeast colonies isolated was 4918. The occurrence of Aspergillus among the filamentous fungi was high, 156 (35%) isolates of positive samples isolated from sputum and lavage.
The most dominant species constituting with the highest frequency were A. niger 41 (9.21%), followed by A. terreus and A. flavus 37 (8.31%) and 34 (7.64%) respectively. Other Aspergilli included A. tubingensis 18 (4.04%), A. nomius 5 (1.12%), and A. fumigatus 4 (0.89%), while non-Aspergillus species included Penicillium as the second one with 151 (33%) isolates and the third one was Cladosporium with 97 (21%) isolates.
Whereas with regard to yeasts, it had relied on its diagnosis on the CHROMO-agar, which showed that 2900 (58.9%) brown yeasts represented the highest one of yeast isolated from lavage, and 833 (16.9%) isolates of Candida parapsilosis, 653 (15.3%) isolates of C. albicans, 250 (5.08%) black yeasts from lavage, as well as 87 (1.76%) C. krusei, 43 (0.87%) C. glabrata, 42 (0.85%) C. dubliniensis, and 10 (0.2%) Rhodotorula sp.
Twenty-three isolates of Aspergillus species of this study were subjected for DNA extraction and PCR assay with AFU5S primer, the molecular weight of PCR product was 236 bp. Sequencing of nitrogen bases for Aspergillus spp. were matched with the reference strains sequence deposited in the Gene Bank using the NCBI BLAST Nucleotide Database program by using AFU5S primer for the 18S region, and the results are identical to diagnose phenotypically for most of the samples under study. This technique is successful in the diagnosis of isolates that did not reach their species by using traditional methods such as A. nomius, E. amstelodami, A. sclerotioniger, and A. tubingensis.
The polymorphism of TLR2 gene was done for 15 samples of blood DNA; the PCR products showed 524 bp. The results of sequences were aligned with the reference sequence in NCBI. There are several genotypes in the TLR2 rs5743708 G/A gene. Each observed substitution or insertion mutation as well as novel SNP (recorded for the first time at this site).
The polymorphism of TLR8 (rs3764880 A/G) gene was the frequency of genotypes AG, AA, and GG respectively 30.0%, 46.6%, and 23.3% in patients. Three genotypes were found in this study of (IL-1RN) gene polymorphism in patients with pulmonary aspergillosis, while there are two genotypes of (IL-1RN) gene polymorphism in the control group.