Detection of Shigella spp. nucleic acids in the synovial tissue of Tunisian rheumatoid arthritis patients and other forms of arthritis by quantitative real-time polymerase chain reaction

• Published in: Rheumatology international Vol. 38; no. 6; pp. 1009 - 1016
• Main Authors: Siala, Mariam, Rihl, Markus, Sellami, Hanen, Znazen, Abir, Sassi, Nadia, Laadhar, Lilia, Gdoura, Radhouane, Belghuith, Imen, Mrabet, Dalila, Baklouti, Sofien, Sellami, Slaheddine, Sibilia, Jean, Fourati, Hela, Hammami, Adnene, Cheour, Ilhem
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-06-2018; Springer Nature B.V
• Subjects: Adult; Arthritis, Rheumatoid - microbiology; Deoxyribonucleic acid; DNA; DNA, Bacterial - analysis; Female; Humans; Infection and Immunity; Male; Medicine; Medicine & Public Health; Middle Aged; Nucleic Acids; Polymerase chain reaction; Real-Time Polymerase Chain Reaction - methods; Reproducibility of Results; Rheumatoid arthritis; Rheumatology; Shigella - isolation & purification; Synovial Membrane - microbiology; Tunisia; Real-time PCR (qPCR); Synovial tissue; spp; Arthritis; Spondyloarthritis; Rheumatoid arthritis; Shigella spp
• ISSN: 0172-8172; 1437-160X
• DOI: 10.1007/s00296-018-3939-y

Enterobacterial components in the joints of patients are believed to contribute to a perpetuating inflammation leading to a reactive arthritis (ReA), a condition in which microbial agents cannot be recovered from the joint. At present, it is unclear whether nucleic acids from
Shigella
spp. are playing a pathogenic role in causing not only ReA but also other forms of arthritis. Quantitative real-time polymerase chain reaction assay (qPCR) is the method of choice for the identification of bacteria within the synovium. The aim of our study was to detect the presence of
Shigella
spp. nucleic acids in the synovial tissue (ST) of Tunisian arthritis patients. We investigated 57 ST samples from rheumatoid arthritis (RA)
n
 = 38, undifferentiated oligoarthritis (UOA)
n
 = 12, and spondyloarthritis (SpA)
n
 = 7 patients; 5 ST samples from healthy individuals were used as controls.
Shigella
spp. DNA and mRNA transcripts encoding the virulence gene A (
VirA
) were examined using an optimized qPCR with newly designed primers and probes. Using qPCR,
Shigella
spp. DNA was found in 37/57 (65%) ST samples (24/38, i.e., 63.2% of RA, 8/12, i.e., 67% of UOA, and 5/7, i.e., 71.4% of SpA patients). Paired DNA and mRNA were extracted from 39 ST samples, whose
VirA
cDNA was found in 29/39 (74.4%) patients. qPCR did not yield any nucleic acids in the five healthy control ST samples. The qPCR assay was sensitive and showed a good intra- and inter-run reproducibility. These preliminary findings generated by an optimized, highly sensitive PCR assay underline a potential role of past gastrointestinal infections. In Tunisian patients, a bacterial etiology involving
Shigella
spp. in the manifestation of arthritic disorders including RA might be more common than expected.