Fluorescence In Situ Hybridization (FISH) and Peptide Nucleic Acid Probe-Based FISH for Diagnosis of Q Fever Endocarditis and Vascular Infections
Authors: Elsa Prudent
Didier Raoult
Hubert Lepidi
Emmanouil Angelakis
Journal: Journal of Clinical Microbiology (2018)
Discipline: MICROBIOLOGY
Keywords: Coxiella burnetii; FISH; PNA probe; Q fever; infective endocarditis
Affiliations: Aix Marseille Université, IRD, AP-HM, MEPHI, IHU-Méditerranée Infection, Marseille, France., Aix Marseille Université, IRD, AP-HM, VITROME, IHU-Méditerranée Infection, Marseille, France., French Reference Center for the Diagnosis and Study of Rickettsioses, Q fever and Bartonelloses, IHU-Méditerranée Infection, Marseille, France., Aix Marseille Université, IRD, AP-HM, MEPHI, IHU-Méditerranée Infection, Marseille, France .

Abstract:  Endocarditis and vascular infections are common manifestations of persistent localized infection due to Coxiella burnetii, and recently, fluorescence in situ hybridization (FISH) was proposed as an alternative tool for their diagnosis. In this study, we evaluated the efficiency of FISH in a series of valve and vascular samples infected by C. burnetii We tested 23 C. burnetii-positive valves and thrombus samples obtained from patients with Q fever endocarditis. Seven aneurysms and thrombus specimens were retrieved from patients with Q fever vascular infections. Samples were analyzed by culture, immunochemistry, and FISH with oligonucleotide and PNA probes targeting C. burnetii-specific 16S rRNA sequences. The immunohistochemical analysis was positive for five (17%) samples with significantly more copies of C. burnetii DNA than the negative ones (P = 0.02). FISH was positive for 13 (43%) samples and presented 43% and 40% sensitivity compared to that for quantitative PCR (qPCR) and culture, respectively. PNA FISH detected C. burnetii in 18 (60%) samples and presented 60% and 55% sensitivity compared to that for qPCR and culture, respectively. Immunohistochemistry had 38% and 28% sensitivity compared to that for FISH and PNA FISH, respectively. Samples found positive by both immunohistochemistry and PNA FISH contained significantly more copies of C. burnetii DNA than the negative ones (P = 0.03). Finally, PNA FISH was more sensitive than FISH (60% versus 43%, respectively) for the detection of C. burnetii We provide evidence that PNA FISH and FISH are important assays for the diagnosis of C. burnetii endocarditis and vascular infections. 

Retracted: No

PubPeer comments:
 This paper mentioned an approval under the IHU ethics committee no. 2016-025. Several others papers, published in different type of journals, mention the same approval number, for different type of test, indications or patients. In this paper, the authors mentioned that no consent has been requested from french participants because it is a retrospective study. According to usual Editors policies, retrospective studies are conducted on already available data or biological material. The Ethics committee nu00b02016-025 seems to have been granted in 2016. However, the authors mentioned that *all these samples were received from both hospitalized patients and 90 outpatients throughout France in our Q fever reference center in Marseille between 2014 and 2017. A study can not be retrospective if the study continue to collect data or materials after the ethics committee approval. In this case, a study should be considered as prospective. Which may also raise an issue concerning the study start date. According Editors guidances and Helsinki Declaration, a prospective study should get before the study start date (recruitment of participants). And participants should give their consents. It seems to be a retrospective ethics committee approval for a prospective interventional study which seriously deviate from Helsinki Declaration. It would be in the public interest if the editor can clarify this point. Otherwise, in France, interventional prospective study MUST get an approval from certified IRB (Comitu00e9 de Protection des Personnes) before to start. The local IHU Ethics Committee is not on the list of the certified IRB and has not the ability to validate a prospective interventional study. It would be in the public interest if the authors can explain why the study did not mention any approval from a Comitu00e9 de Protection des Personnes before to start the recruitment. We can also note that several other papers mentioned the same ethics approval number. Usually, one ethical approval number is attached to one protocol. In the other paper, we can note : differents type of bacteria, patients in another country for a different indication (tuberculosis), patient with tobacco issue performing CT scan... It should be in the public interest to understand how a same ethical approval number for a protocol can cover so many different type of situations. - [Extended spectrum of antibiotic susceptibility for tuberculosis, Djibouti] - [u201cMycobacterium mephinesiau201d, a Mycobacterium terrae complex species of clinical interest isolated in French Polynesia] - [Pulmonary Isolation of Multidrug resistant u201cMycobacterium simulansu201d and Mycobacterium tuberculosis from a patient in the Horn of Africa] - [Fluorescence in situ hybridization, a complementary molecular tool for the clinical diagnosis of infectious diseases by intracellular and fastidious bacteria] - [Fluorescent in situ hybridization can be used as a complementary assay for the diagnosis of Tropheryma whipplei infection] , 
 Expression of Concern, 7 September 2022: https: journals.asm.org doi 10.1128 jcm.01077-22 It reads:  *The American Society for Microbiology was notified that the above-mentioned paper is being reviewed as part of a u201cscientific misconduct investigationu201d by the University of Aix Marseille. This Expression of Concern is issued pending the outcome of the investigation and will be updated accordingly thereafter.* ,