Urinary infections are a serious health problem, in part because of their prevalence. Clinical and experimental data confirm that the advancement of microorganisms along the urethra is the most frequent route of urinary infection.

Pathways, especially for organisms derived from the intestine (i.e., Escherichia coli and other Enterobacteriaceae.

This also provides a logical explanation for the higher frequency of MI in women than in men and an increased risk of infection after catheterization [1,2]. Escherichia coli can be defined as a rod-shaped bacterium that lives in the intestines of humans and some animals. There are more than a hundred types of this stick. Most of them are completely harmless, but some can cause serious illness [3,4].

Some serious strains of E. coli can cause hemolytic uremic syndrome (HUS). This is a dangerous condition in which a large number of blood clots form in small vessels, erythrocytes are damaged, and many organ failures occur - a severe stress reaction of the body with any suspicion of HUS, an analysis for (dangerous) enterosorbents is required [5-7]. And usual bacteriological analysis, which is carried out in inflammatory diseases of the reproductive system, postpartum endometritis, and others, can show the presence of E. coli [8].

E. coli bacteria belong to the Enterobacteriaceae family, whose members are ubiquitous. They are widespread in soil, water, and plants. They are part of the intestinal flora of most animals, including humans. This family causes a variety of human diseases, including bacteremia, infections, Urinary tract (UTI), and many intestinal infections. E. coli is characterized by short Gram-negative bacilli that do not form facultatively aerobic or anaerobic sporophytes and grow in simple culture media. The optimum temperature for growth is 37 °C (Gupte), 2010) [9-11]. And they are circular, smooth, and convex colonies with specific edges and non-mucosal, some of their strains produce hemolysin on the medium (Blood agar). The color of the colony on the Maconque medium is pink, but on the medium (EMB agar), it is characterized by green metallic shine, movable by peripheral flagella or non-motile, and negative for the oxidase test [12,13]. Positive for the indole and methyl red test  and negative for the Fuchsprosecure and citrate test, and it ferments the sugars lactose, glucose, sucrose, maltose, and mannitol and produces acid and gas, and does not decompose urease and does not produce gas Gupte, [14] (HQS). As for the antigenic properties, these bacteria contain three types of antigens. O antigen (Somatic antigen), K antigen (Casular antigen), and flagellated antigen Brook 2004) (H antigen) Flagellar antigen. E. coli is a pathogen.

The most common opportunists worldwide are involved in microbial contamination and cause about 90% of the alternating current [14].  

Problem Statement

Urinary tract infection is one of the most common problems in primary health care and the majority of the community. Urinary tract infection is among the most common and highly morbidity infectious diseases. Moreover, the attractiveness is presented in a diverse way; Cystitis, pyelonephritis, septic shock, to multi-organ failure [2].

Escherichia coli is a common causative agent Urinary tract infections cause between 70 and 95% of urinary tract infections [8]. Strains Escherichia coli, the urogenital pathogen responsible for 69% of associated cases of Cystitis, 67% of cases of pyelonephritis, and 72% of cases associated with this research aims to find out the determination in Intestinal and Urinary tract infection by detection molecular for gene hlyA.

UPEC strains can be found in the human gastrointestinal tract. From there, it can reach the urinary tract and cause an infection using a set of multiple virulence genes to colonize and infect upward through the ureters, which can cause damage to kidney or even sepsis. In some cases, the spread of a single clonal group of UPEC strains can occur within a community through contaminated food or through pets.

UTIs caused by E. coli are ascending infections in which bacterial contamination of the periurethral space allows UPEC access to the urinary tract. In general, the rise of UPEC is mediated by the action of flagella, which drive bacteria from the urethra to the bladder, which in turn is colonized through fimbrial and afimbrial adhesions; they use iron uptake systems to facilitate their growth as well as toxins to avoid the innate immune response. Flagella also mediate the ascension of UPEC to the kidneys, so motility, flagellar, is essential for uropathogenesis, and fimbrial and afimbrial adhesions allow UPEC to support the flow of urine that can carry bacteria out of the urinary tract [15].

More specifically, UPEC entry into the urinary tract is followed by adherence to the uroepithelium, mediated by fimbrial adhesin H that binds to integrins α3 and β1, which group with actin at invasion sites, thus as destabilization of microtubules, resulting in the internalization of the bacteria Once internalized, UPEC rapidly replicates and forms microbial communities or biofilms, called intracellular bacterial communities (CBIs) that serve as protective environments. UPEC can leave the CBIs and enter the bladder lumen, where the infection promotes the influx of polymorphonuclear cells, causing tissue damage, apoptosis, and exfoliation of bladder cells. At this point in the infection, if not treated properly, it can ascend to the kidneys, causing pyelonephritis. 

Patient And Method

Patient Simple: In this study, 218 samples were collected from Yarmouk Hospital, Iraq, Baghdad, and The study was specialized in urological patients and the identification of the hlyA gene by relying on molecular detection.

Study Design

In this study, a gene hlyA is partially investigated and detected by relying on the PCR assay, and the design was also based on the NCBI-Gen Bank published sequence, Escherichia coli strain BAU-MH4 HlyA (hlyA) gene, partial CDs.

Urinary tract infections belong to the most common infectious diseases in the world; they represent the highest incidence and prevalence of E. coli Urinary pathogen is the extra-intestinal pathogen that is frequently isolated from UTI patients and is the cause of 75 to 95% of all cases. In Iraq, Escherichia coli is one of the pathogens that causes UTIs and is more common in women, with a higher incidence and prevalence

Study Period

Samples were collected from the hospital and the study period was 9-9-2017 to 12-12-2019.

Aim Of Research 

The research aims to know and evaluate the determination in Intestinal and Urinary tract infection by detection molecular for gene hlyA.

Ethics Statement

The ethics committee Institutional approved the study, identity of the participants. They were kept in reserve, and the results were delivered individually electronically.

Two hundred eighteen samples were collected from Yarmouk Hospital, Iraq, Baghdad, where the research aims to Find out the determination in Intestinal and Urinary tract infection by detection molecular for gene hlyA (Table 1). 

Through Table 2, it becomes clear to us that culture growth distribution for urine.

The positive group was 112 samples, while the negative group was 60 samples, and the ages were limited to between 8 and 17 years; and by relying on statistical analysis to analyze data and demographic information, the mean Sd was found to reach the age of 11.78±2.9 as shown in Tables 2,3,4.

Through Table 5, we can see the distribution according to gender, where the percentage of growth culture positive for females was 65.17% (73) speciman, but for males, it was 34% (39) speciman.

As for the diarrhea, samples were 150, distributed among 120 samples: growth culture-positive either, 30 negative, and distributed growth culture-positive 85 samples for males and 35 samples for females. As for Detection of hemolysin, E.coli was 40 samples of urine and 33 samples of diarrea. Molecular characteristic of the hlyA gene shows that 11 isolates were produced from E.coli and divided into eight isolated from urine and three from diarrea (Table 6,7). Although appropriate use of antibiotics is the current focus in Iraq, there is little information published about the impact in the country regarding inappropriate use of antibiotics in terms of bacterial resistance. Currently, there is no comprehensive surveillance for community-acquired UTIs in our country, so it is very difficult to estimate the true incidence of UTIs and the potential treatment that we can use to control the spread of these organisms (Table 8). On the other hand, the formation of biofilms (microbial communities and/or aggregates adhered to an extracellular matrix that they produce themselves) is today considered a potential risk to human health, given that it is associated with antimicrobial resistance and needs to be located for appropriate treatment for related chronic infections. At present, several strategies have been proposed with the aim of preventing the formation of these microbial aggregates by different substances in both solution and their application to medical devices (Table 9).

Table 1: Detection of Hemolysin

Table 2: Culture Growth Distribution Urine

Table 3: Growth Positive Distributed Depend on Age

Table 4: Mean SD (Age)

Table 5: Growth Positive Distributed Depend on Age

Table 6: Diarrhea Samples

Table 7: Distribution of Diarrhea Samples Depends on Gender (+)

Table 8: Detection of Hemolysin Escherichia Coli (S Test).

Table 9: Optimal Conditions for Gene Diagnosis

This study concludes that the presence of the gene in E. coli bacteria and the percentage was not normal. On the contrary, the gene was present in a very high percentage. Our study was supportive and proves the validity of other studies, such as the study of Dr. Kafl, who concluded in his study that the hylA gene was found in a percentage of 90% in E. coli bacteria. His study was also based on molecular detection.

Recommendation

• Conducting studies dealing with the link between the effect of antibiotics and virulence factors in bacteria that cause urinary tract infections and its relationship to human health

• Studying the patterns of hemolysis in humans and other mammals and revealing the different mechanisms of the effect of antibiotics and antiseptics on the properties of bacteria and their ability to decompose blood, which participate in the development of the work of these substances in affecting pathogenic bacteria in the future

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