Clinical trial NCT05229822: Bacterial Translocation Markers as Predictors of Infectious and Inflammatory Complications in Acute Bowel Obstruction
Official title: Prognostic Significance of Bacterial Translocation Markers as Predictors of Infectious and Inflammatory Complications in Acute Mechanical Bowel Obstruction
Study type: OBSERVATIONAL
Overall status: COMPLETED
Conditions: Intestinal Obstruction
Colorectal Cancer
Post-Op Complication
Systemic Inflammatory Response Syndrome
Colon Tumor
Keywords: acute bowel obstruction
colon tumor
bacterial translocation
presepsin
lipopolysaccharide-binding protein
16s rRNA
post-operative complications
Lead sponsor: Karaganda Medical University
Sponsor class: OTHER
Collaborators: name: Ministry of Education and Science, Republic of Kazakhstan
First posted: 2022-02-08
Enrollment: 150
Sex: ALL
Minimum age (years): 18
Standard ages: ADULT
OLDER_ADULT
Accepts healthy volunteers: no
Observational model: OTHER
Time perspective: PROSPECTIVE
Sampling method: PROBABILITY_SAMPLE
Study population: Patients with malignant and benign acute bowel obstruction, colorectal cancer patients without acute bowel obstruction (planned operations) aged from 18, who are expected to undergo surgical treatment for this pathology.

Brief summary: Despite modern approaches to the diagnosis and treatment of acute bowel obstruction (ABO), postoperative mortality ranges from 5 to 32%, and complications occur up 23% of cases. One of the formidable infectious and inflammatory complications of ABO is sepsis. The main component of the development of sepsis in ABO is bacterial translocation (BT). BT is the migration of intestinal bacteria or their products through the intestinal mucosa into the mesenteric lymph nodes and further into normally sterile tissues and organs.

Today there are several methods for detecting BT:

1. direct method - the detection of 16s rRNA (ribosomal ribonucleic acid) in mesenteric lymph nodes (MLN);
2. indirect method - the detection of serum lipopolysaccharide-binding protein (LBP) and presepsin (Soluble CD14 subtype or sCD14-ST).

The aim of this study is to determine the diagnostic and prognostic significance of bacterial translocation as a predictor of the complications development in patients with malignant and benign acute bowel obstruction by assessing the relationship of biomarkers in the systemic circulation (LBP, sCD14-ST) with the detection of microorganism genes (16s rRNA) in mesenteric lymph nodes.

Detailed description: For the early diagnosis of infectious and inflammatory complications, it is necessary to study LBP, sCD-14 and 16sRNA as bacterial translocation markers in patients with malignant and benign acute bowel obstruction, as well as in patients after planned surgical intervention for colon tumors. Based on changes in bacterial translocation biomarkers in the blood serum, it's suggested that patients with researched pathology can be stratified according to the risk level of developing infectious and inflammatory complications.

The study materials are blood serum and mesenteric lymph nodes (MLN). Venous blood sampling will be performed 1 hour before surgery, 24 and 72 hours after it. Venous blood will be collected in 5 ml vacutainers with a coagulation activator and a serum gel separator. It will be centrifuged for 20 minutes at 1000 x g, after which the gel completely separates the serum from the clot, forming a tight barrier.ELISA Kit for Lipopolysaccharide Binding Protein (LBP, Human) and for Presepsin (sCD14-ST, Human), from Cloud-Clone Corp. will be used to determine any presence of LBP and sCD14-ST. The analysis will be performed according to the manufacturer's instructions for an ELISA EVOLIS robotic system from BioRad.

The operating surgeon will perform a MLN sampling in sterile conditions during surgery after resection of the intestine from the mesentery of the gross specimen. MLN will be placed in a sterile tube without any fillers. The DNA will be extracted by the GeneJET Genomic DNA Purification Kit manufactured by Thermo Fisher Scientific, USA, in accordance with the manufacturer's instructions. The 16s rRNA bacteria in MLN will be detected by using real-time PCR and BIO-RAD CFX96 amplifier with 16s rRNA forward and reverse primers (U16SRT-F FACTCCTACGGGAGGGAGGCAGGT and U16SRT-R TATTACCGCGGCTGCTGGGC).

During the implementation, the resources of the Collective Use Laboratory of Research Center Non-profit Joint Stock Company (NJSC) "Karaganda Medical University" will be used.

This research is funded by the Science Committee of the Ministry of Education and Science of the Republic of Kazakhstan (Grant No. AP09260597).

Eligibility criteria: Inclusion Criteria:

* patients with malignant acute bowel obstruction,
* patients with benign acute bowel obstruction,
* colorectal cancer patients without acute bowel obstruction (planned operations).

Exclusion Criteria:

* age less than 18,
* pregnancy,
* patients with paralytic acute bowel obstruction,
* patients with HIV infection, liver cirrhosis,
* patient with an infectious process due to another pathology.

Primary outcomes:
measure: Change in Number of Participants With Post-operative Infectious and Inflammatory Complications
timeFrame: Once (if any complication occurs during hospitalization - from 7 to 28 days)
Secondary outcomes:
measure: LBP Level in Serum Blood
timeFrame: 1 hour before surgery, 72 hours after surgery
measure: sCD14-ST Level in Serum Blood
timeFrame: 1 hour before surgery, 72 hours after surgery
measure: 16s rRNA in Mesenteric Lymph Nodes
timeFrame: Once (MLN sampling in sterile conditions during surgery)
Other outcomes:
Locations:
city: Karaganda
country: Kazakhstan
facility: NJSC Karaganda Medical University
Study officials:
affiliation: NJSC Karaganda Medical University
name: Yemek Turgunov, Pr.
role: STUDY_CHAIR
affiliation: NJSC Karaganda Medical University
name: Alina Ogizbayeva, PhD student
role: PRINCIPAL_INVESTIGATOR
affiliation: NJSC Karaganda Medical University
name: Lyudmila Akhmaltdinova, PhD
role: PRINCIPAL_INVESTIGATOR
affiliation: NJSC Karaganda Medical University
name: Kairat Shakeyev, Pr.
role: PRINCIPAL_INVESTIGATOR
affiliation: Multidisciplinary hospital No. 1 of Karaganda
name: Dmitry Matyushko, PhD
role: PRINCIPAL_INVESTIGATOR
affiliation: NJSC Karaganda Medical University
name: Miras Mugazov, PhD
role: PRINCIPAL_INVESTIGATOR
affiliation: Multidisciplinary hospital No. 3 of Karaganda
name: Asylbek Zhumakaev, Master
role: PRINCIPAL_INVESTIGATOR
affiliation: NJSC Karaganda Medical University
name: Irina Kadyrova, PhD
role: PRINCIPAL_INVESTIGATOR
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