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The National Reference Laboratory (NRL) for Escherichia coli, particularly referring to E. coli strains pathogenic to humans, hosted at the Department of Microbiological Food Safety and Foodborne Diseases, in the Department of Food Safety, Nutrition and Veterinary Public Health (DSANV) of the Istituto Superiore di Sanità (ISS).

The laboratory was designated as NRL by the Ministry of Health with communication dated 14 November 2006, for application of the art. 33 of Regulation (EC) 882/2004.

The Ministry of Health has also designated the ISS as the Reference Laboratory for the diagnosis and surveillance of STEC infections in humans and as the Italian Operational Contact Point (OCP) of the European Center for Disease Control (ECDC) for these infections.

The NRL E. coli also carries out the functions of the European Reference Laboratory for E. coli (EU Reference Laboratory for E. coli, EURL-VTEC) by mandate of the European Commission, in order to provide qualified and recognized performances and results in the international field.

Both the NRL E. coli and the EURL-VTEC operate in compliance with the UNI CEI EN ISO/IEC 17025 standard within the DSANV Quality Management System (accreditation No. 0779 L), with both fixed and flexible scope of accreditation.

Diagnostic activity and methods
The diagnostic activities performed at the NRL for E. coli include:

STEC detection in food samples

This is carried out by applying the internal method LNRVTE53, consisting in the following procedures:

Horizontal method for the detection of Shiga toxin producing Escherichia coli (STEC) and the determination of O157, O111, O26, O103 and O145 serogroups – ISO TS 13136:2012

Detection and identification of Shiga toxin-producing Escherichia coli (STEC) O104:H4 in food by Real Time PCR – EURL-VTEC Method 04_Rev 2

Detection of Escherichia coli producing the Stx2f subtype by Real-Time PCR – EURL-VTEC Method 10_Rev 0.

STEC detection in human/animal clinical samples (faeces or other biological materials)

These tests are carried out using the LNRVTEC29.0n accredited internal method (screening for the presence of virulence genes and genes associated with the main STEC serogroups through RT-PCR) and the LNRVTEC09.0n accredited internal method (isolation of STEC strains from samples resulted positive to the screening step):

identification of the STEC strains serogroup (LNRVTEC08.0n internal accredited method)
detection of free Shiga toxin (Stx) in human stool extract (cytotoxicity assay on VERO cell monolayer. LNRVTEC07 Internal method - Non accredited)
detection and typing of STEC strains (by confirming the presence of virulence genes using the LNRVTEC09.0n accredited internal method)
serological diagnosis of STEC infection in humans (ELISA method to detect circulating antibodies direct against lipo-polysaccharide (LPS) of the serogroups most associated with human infections: E. coli O157, O26, O103, O111, O145. LNRVTEC06 Internal method - Non accredited)

Diagnostic services can be requested by using the "Analysis request form (PGCPSP01.E0n)", attached to this page. The form also contains the analytical details of each test and instructions for sending the samples.