تغاير العلاقات التطورية وتصويف عوامل الضراوة لعزلات المكورات العنقودية من المرضى في محافظة بابل == Phylogenetic diversity and characterization of virulence factor for Staphylococcus aureus isolated from patients in Babylon province
Author name:
نبراس عودة كاظم عيسى
Supervisor name:
لميس عبد الرزاق عبد اللطيف
General topic:
Medicine
Specific topic:
Microbiology
Degree:
Master
University:
University of Babylon - Faculty Of Medicine
Language:
English
University location:
Babylon
First pages:
19T1274 - p.pdf
Abstract:
A total 100 samples, only nineteen isolates of S. aureus were obtained from different samples includes (urine ,vagina, stool, wound, and abscess ) from patients by standard bacteriological methods. All samples were obtained from patients or individuals who admitted to Al - Hillah Surgical Teaching Hospital and Merjan Medical City in Babylon Governorate during a period extending from (July 2017 to September 2017). Then the specimens cultured in selective media and identified by using biochemical and bacteriological tests and using specific primer(sau).Only (19)isolate of Staphylococcus aureus isolates were recovered.Some virulence factors of all S. aureus isolates were studied as sea gene , PCR was performed using specific primer on genomic DNA from S .aureus isolate . PCR amplification of sea gene indicated that two from vagina isolate 2 : 4(50%),three from urine 3 : 3(100%) and four from stool 4 : 4(100%) and one isolates from abscess 1 : 4(25%) and one isolates from wound1 : 4(25%) .Specific PCR primer was used for the detection of( tsst) gene.It was found that tsst gene was observed in 16 isolates which include urine, pus ,wound,and stool in percentage 100%.Specific primer was used for detection of eta gene .It was found that eta was observed in three from urine 3 : 3(100%), four from vagina 4 : 4(100%) and three from abscess 3 : 4(75%) and four from wound4 : 4(100%) and four from stool 4 : 4(100%).cna gene was also detected in S.aureus by using specific PCR primer .It was found that can gene was observed in two from urine 2 : 3(66.6), one from vaginaII1 : 4(25%), three from wound 3 : 4(75%) and three from abscess 3 : 4(75%) and two from stool 2 : 4(50%) .Also, ica gene was detected in S.aureus isolates by using specific primer , it was found that gene present in 2(66.6%) isolate from urine sample and 1(25%) isolate from abscess sample.However, molecular detection of pvl gene was done using specific PCR primer. It was found that pvl was observed in bacterial isolates in only 2(50%) isolate from abscess sample and 1(25%) from wound sample.Clumping factor A (clfA) gene was investigated by PCR technique using specific primer for his gene. It was found that clfA gene was observed in three from urine sample 3(100%) and one from vagina 1(25%) and four from abscess 4(100%) and four from wound 4(100%) and three from stool 3(75%).S. aureus isolates were subjected to show its ability to produce staphylokinase enzyme .It was that only 2 from 4 (50%) isolates from wound extracylallary were able to produce staphylokinase enzyme by creating clear zones around wells containing their cultures in the skim milk agar plate.Hyaluronidase activity was also investigated it was found that( 89%) of S. aureus isolates give positive result for hyaluronidase by turbidity reduction assay in the determination of hyaluronidase.The relationship between 19 S. aureus isolates using RAPD - PCR was performed. Ten primer showed polymorphism among the isolates test generatingIII88 bands, 58 of which were polymorphic with sizes ranging between 100 to 3000 bp. The mean percentage of ten primer 61.36% .The Dendogram two main cluster, the first include one isolate (st9 abscess) and the second include 18 isolates, However , the second cluster divide the 18 isolate into two subcluster. Using the ten primer isolate 11 reveled highest similarity with isolate 1 (69%) and the isolate 17 and 18 have similarity (69%) followed by isolates 7 and 16 with similarty (61%) .However , isolate 14 and 9 showed the lowest similarity (10%). All the remaining pairs showed intermediate similarity percentage with highest and lowest percentage .PCR amplification of Agr group gene from 19 isolates product different types based on size range from (323 - 659 ) bp were recognized as follows Agr I (441bP) ,Agr II(575 bp),Agr III(323 bP),Agr IV (659 bp),and. The frequencies of agr Groups III 17(89.1%) and Agr Groups I 15(78.7%) were more than those seen in other Groups flowed by agr IV 7(36.7%), while agr Group II 2(10.4%) was the least obtained in all the five isolates .DNA sequencing of cna gene from isolates of S. aureus revealed 188 mutation in four isolates and give identity in percentage (76 - 89%) with standard strand S. aureus in NCBI website ,this gene sequencing is the first described in Iraq.
