Novel diagnostic approach for amoebic liver abscess using cell free (cf) DNA: a prospective study

Abstract

Background

Amoebic liver abscess (ALA) is commonly seen in tropical countries and diagnosis of ALA relies mainly on non-specific serological and imaging techniques as well as PCR from pus.

Objective

This study evaluated the potential of using cell free DNA (cfDNA) from serum and urine for diagnosing ALA.

Methods

We prospectively evaluated quantitative PCR (qPCR) for detection of cf DNA in serum and urine sample in all liver abscess patients. The samples were collected from patients reporting to emergency ward of Postgraduate Institute of Medical Education and Research, Chandigarh, India with symptoms suggestive of liver abscess. Real time PCR was done to detect cf DNA in serum and urine by targeting 99-bp unit of small subunit rRNA of Entamoeba histolytica and conventional PCR for pus.

Results

A total 113 samples (serum and urine) and 100 pus samples were analysed. A total of 62 ALA patients were confirmed; with maximum 57 patients detected by qPCR for cfDNA in the serum, 55 patients by PCR on pus aspirate and 50 ALA patients by qPCR for cfDNA in urine sample. Therefore, the sensitivity of qPCR for detection of cf DNA in serum was 91.94% and for urine was 80.65%.

Conclusion

A total of 11.2% of ALA patients were diagnosed only through detection of E. histolytica cf DNA in their serum and urine. Detection of cfDNA from serum, urine of ALA has a potential role in future especially for developing countries as it is a rapid, sensitive and patient friendly diagnostic approach.

Keywords:

• Amoebic liver abscess
• cell free DNA
• urine
• serum
• real-time PCR

Author contributions

Priya Datta: Revised the manuscript, contributed to the conception of the study, and gave final approval of the version to be submitted. Divya Rattan: was involved in the acquisition of data, analysed, and interpreted the data. Devyani Sharma: Sample collection & processing. Navneet Sharma: Revised the manuscript, contributed to the conception of the study and provide samples. Naveen Kalra: Revised the manuscript and provide samples. Ajay Duseja: Revised the manuscript and provide samples. Archana Angrup: Data analysis. Rakesh Sehgal: Revised the manuscript.

Ethics approval and consent to participate

This study was approved by Institutional Ethics Committee of Postgraduate Institute of Medical Education and Research, Chandigarh, India vide no. PGI/IEC/2022/000748 and the consent from the participants were taken on the consent performa.

Disclosure statement

No potential conflict of interest was reported by the authors.

Availability of data and materials

The data produced from the current study is available online on NCBI database.

Additional information

Funding

This study was funded by ICMR funded project (Sanction order no. OMI/11/2022/ECD, PD).