Bacterial Translocation Markers as Predictors of Infectious and Inflammatory Complications in Acute Bowel Obstruction

NCT ID: NCT05229822

Last Updated: 2025-01-16

Study Results

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Basic Information

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Recruitment Status

COMPLETED

Total Enrollment

150 participants

Study Classification

OBSERVATIONAL

Study Start Date

2021-03-01

Study Completion Date

2023-12-31

Brief Summary

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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

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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).

Conditions

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Intestinal Obstruction Colorectal Cancer Post-Op Complication Systemic Inflammatory Response Syndrome Colon Tumor

Study Design

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Observational Model Type

OTHER

Study Time Perspective

PROSPECTIVE

Study Groups

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Malignant ABO

60 patients with malignant acute bowel obstruction

LBP

Intervention Type DIAGNOSTIC_TEST

Determine any presence of LBP in blood serum by ELISA method 1 hour before surgery, 24 and 72 hours after it.

sCD14-ST

Intervention Type DIAGNOSTIC_TEST

Determine any presence of sCD14-ST in blood serum by ELISA method 1 hour before surgery, 24 and 72 hours after it.

16s rRNA

Intervention Type DIAGNOSTIC_TEST

Determine any presence of 16s rRNA in mesenteric lymph nodes by PCR method.

CRC without ABO (control)

60 colorectal cancer patients without acute bowel obstruction (planned operations)

LBP

Intervention Type DIAGNOSTIC_TEST

Determine any presence of LBP in blood serum by ELISA method 1 hour before surgery, 24 and 72 hours after it.

sCD14-ST

Intervention Type DIAGNOSTIC_TEST

Determine any presence of sCD14-ST in blood serum by ELISA method 1 hour before surgery, 24 and 72 hours after it.

16s rRNA

Intervention Type DIAGNOSTIC_TEST

Determine any presence of 16s rRNA in mesenteric lymph nodes by PCR method.

Benign ABO

30 patients with benign acute bowel obstruction

LBP

Intervention Type DIAGNOSTIC_TEST

Determine any presence of LBP in blood serum by ELISA method 1 hour before surgery, 24 and 72 hours after it.

sCD14-ST

Intervention Type DIAGNOSTIC_TEST

Determine any presence of sCD14-ST in blood serum by ELISA method 1 hour before surgery, 24 and 72 hours after it.

16s rRNA

Intervention Type DIAGNOSTIC_TEST

Determine any presence of 16s rRNA in mesenteric lymph nodes by PCR method.

Interventions

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LBP

Determine any presence of LBP in blood serum by ELISA method 1 hour before surgery, 24 and 72 hours after it.

Intervention Type DIAGNOSTIC_TEST

sCD14-ST

Determine any presence of sCD14-ST in blood serum by ELISA method 1 hour before surgery, 24 and 72 hours after it.

Intervention Type DIAGNOSTIC_TEST

16s rRNA

Determine any presence of 16s rRNA in mesenteric lymph nodes by PCR method.

Intervention Type DIAGNOSTIC_TEST

Eligibility Criteria

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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.
Minimum Eligible Age

18 Years

Eligible Sex

ALL

Accepts Healthy Volunteers

No

Sponsors

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Ministry of Education and Science, Republic of Kazakhstan

OTHER_GOV

Sponsor Role collaborator

Karaganda Medical University

OTHER

Sponsor Role lead

Responsible Party

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Responsibility Role SPONSOR

Principal Investigators

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Yemek Turgunov, Pr.

Role: STUDY_CHAIR

NJSC Karaganda Medical University

Alina Ogizbayeva, PhD student

Role: PRINCIPAL_INVESTIGATOR

NJSC Karaganda Medical University

Lyudmila Akhmaltdinova, PhD

Role: PRINCIPAL_INVESTIGATOR

NJSC Karaganda Medical University

Kairat Shakeyev, Pr.

Role: PRINCIPAL_INVESTIGATOR

NJSC Karaganda Medical University

Dmitry Matyushko, PhD

Role: PRINCIPAL_INVESTIGATOR

Multidisciplinary hospital No. 1 of Karaganda

Miras Mugazov, PhD

Role: PRINCIPAL_INVESTIGATOR

NJSC Karaganda Medical University

Asylbek Zhumakaev, Master

Role: PRINCIPAL_INVESTIGATOR

Multidisciplinary hospital No. 3 of Karaganda

Irina Kadyrova, PhD

Role: PRINCIPAL_INVESTIGATOR

NJSC Karaganda Medical University

Locations

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NJSC Karaganda Medical University

Karaganda, , Kazakhstan

Site Status

Countries

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Kazakhstan

References

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Gore RM, Silvers RI, Thakrar KH, Wenzke DR, Mehta UK, Newmark GM, Berlin JW. Bowel Obstruction. Radiol Clin North Am. 2015 Nov;53(6):1225-40. doi: 10.1016/j.rcl.2015.06.008.

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Roses RE, Folkert IW, Krouse RS. Malignant Bowel Obstruction: Reappraising the Value of Surgery. Surg Oncol Clin N Am. 2018 Oct;27(4):705-715. doi: 10.1016/j.soc.2018.05.010. Epub 2018 Jul 21.

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Reference Type BACKGROUND
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MacFie J, Reddy BS, Gatt M, Jain PK, Sowdi R, Mitchell CJ. Bacterial translocation studied in 927 patients over 13 years. Br J Surg. 2006 Jan;93(1):87-93. doi: 10.1002/bjs.5184.

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Fang L, Xu Z, Wang GS, Ji FY, Mei CX, Liu J, Wu GM. Directed evolution of an LBP/CD14 inhibitory peptide and its anti-endotoxin activity. PLoS One. 2014 Jul 15;9(7):e101406. doi: 10.1371/journal.pone.0101406. eCollection 2014.

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Mussap M, Noto A, Fravega M, Fanos V. Soluble CD14 subtype presepsin (sCD14-ST) and lipopolysaccharide binding protein (LBP) in neonatal sepsis: new clinical and analytical perspectives for two old biomarkers. J Matern Fetal Neonatal Med. 2011 Oct;24 Suppl 2:12-4. doi: 10.3109/14767058.2011.601923.

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Hosomi S, Yamagami H, Itani S, Yukawa T, Otani K, Nagami Y, Tanaka F, Taira K, Kamata N, Tanigawa T, Shiba M, Watanabe T, Fujiwara Y. Sepsis Markers Soluble IL-2 Receptor and Soluble CD14 Subtype as Potential Biomarkers for Complete Mucosal Healing in Patients With Inflammatory Bowel Disease. J Crohns Colitis. 2018 Jan 5;12(1):87-95. doi: 10.1093/ecco-jcc/jjx124.

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Provided Documents

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Document Type: Study Protocol and Statistical Analysis Plan

View Document

Related Links

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http://doi.org/10.1016/j.clinbiochem.2011.01.012

Mierzchala M, Krzystek-Korpacka M, Gamian A, Durek G. Quantitative indices of dynamics in concentrations of lipopolysaccharide-binding protein (LBP) as prognostic factors in severe sepsis/septic shock patients - Comparison with CRP and procalcitonin /

https://doi.org/10.1007/s00134-014-3514-2

Masson S, Caironi P, Fanizza C, Thomae R, Bernasconi R, Noto A, Oggioni R, Pasetti GS, Romero M, Tognoni G, Latini R, Gattinoni L. Circulating presepsin (soluble CD14 subtype) as a marker of host response in patients with severe sepsis or septic shock

Other Identifiers

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BT-ABO

Identifier Type: -

Identifier Source: org_study_id

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