Tag Archives: C. diff. PCR

C. difficile Laboratory Test Information For Patients and Healthcare Providers

Testing for Clostridium difficile toxin with real-time polymerase chain reaction (PCR) can improve the laboratory diagnoses of C difficile–associated diarrhea, compared with A/B enzyme immunoassay (EIA), according to results from a study presented at a Association for Molecular Pathology meeting.

In fact, real-time PCR demonstrated a sensitivity of 100% and a specificity of 96.9%, whereas the A/B EIA was found to generate both false-positive and false-negative results.

“Although C difficile–associated diarrhea has increased in prevalence and severity, the inaccuracy of conventional C difficile toxin EIA for diagnosis is well documented in the literature,” said lead investigator Cynthia Essmyer, MD, medical pathologist at Saint Luke’s Hospital in Kansas City, Missouri. (1)

The Microbiology Laboratory medical pathologist at a University Hospital located in NC shared the following information to answer one of the many questions:  “Why won’t the lab run a test on hard formed stool?”

The lab will not accept a stool specimen that is hard/formed stool due to the fact that the PCR test is regulated by the testing company and a hard/formed stool can produce a false positive result.  Soft, unformed or watery stools are all accepted for the PCR testing.

  • ACCEPTABLE SPECIMEN: FecesCollect specimen as follows:
    1. Collect feces in a clean, dry container or bedpan not contaminated with urine, residual soap, or disinfectants.
    2. Transfer appropriate volume of feces to a clean dry tightly capped specimen cup.
    3. Submit double-bagged specimen cup immediately to laboratory.
  • NOTES:
    1. Patients should be passing at least 3 unformed or watery stool specimens in a 24-hour period. Most patients have more than 3 episodes of watery unformed stools per day.
    2. Soft specimen is defined as specimen assuming the shape of its container (unformed).
    3. Formed or hard fecal specimens will not be tested.
    4. Repeat testing following a negative test during the same episode of diarrhea is NOT recommended for at least 7-days because of high sensitivity (between 98-99% and with a 99% negative predictive value).

* There may be an alternate send out micro. test for hard formed stool and this information can be obtained at the laboratory being utilized.  Have a healthcare provider contact the lab prior to delivering a specimen to the lab location.

When a PCR result is NEGATIVE: the Wait time between testing is 7 days to retest.

When a PCR result is POSITIVE: the Wait time between testing is 14 days to retest.

Q:  Is there a wait time after completing antibiotic treatment (greater than 14 days since first positive result) to retest?
A:  No, since the PCR is testing micro-organisms the antibiotic will not interfere with testing.


Q:  Can a stool specimen be kept in a refrigerator and if so for how long?

REFRIGERATE IF TRANSPORT IS DELAYED. Specimens can be stored at 2-8 degrees C for 24 hours before significant degredation of the toxin is noted. SUBMIT WITH COLD PACKS OR ON ICE IF TRANSPORT WILL TAKE > 1 HOUR.

A specimen can be rejected by the lab if Specimen is received > 2 hours after collection unless submitted on ice or cold pack or with note that specimen stored in refrigerator prior to transport.

Q:  How long does it take to receive lab results?

A:  Routine turn around time is one day.



Source: (1) Medscape

other:  University Hospital Microbiology Laboratory in NC

ROCHE cobas® C. diff. Test approved by US Food and Drug Administration (FDA)

US Food and Drug Administration (FDA) has provided 510(k) clearance for the cobas® Cdiff Test to detect Clostridium difficile (C. difficile) in stool specimens.

The cobas® Cdiff Test targets the toxin B gene found in toxigenic C. difficile strains directly in specimens from symptomatic patients. The test provides accurate information which assists clinicians in making timely treatment decisions and aids in the prevention of further infection in healthcare settings.

“Having the ability to provide a result quickly is important when supporting infection control for Clostridium difficile,” said Dr. Steve Young, Professor of Pathology, Department of Pathology UNMHSC and Tricore Reference Lab. “The cobas® 4800 System has the capability to allow for mixed batch testing of the cobas® Cdiff Test alongside testing for Methicillin-resistant Staphylococcus aureus, Staphylococcus aureus, and herpes simplex virus 1 and 2*, all on one platform. We can run these assays together at least once in each shift rather than once a day, which can greatly improve laboratory efficiency, ultimately leading to better infection control and patient care.”

In a clinical trial program conducted at sites throughout the United States, the cobas® Cdiff Test demonstrated excellent performance compared to direct and enrichment toxigenic culture. The test combines high assay sensitivity with rapid turnaround time and a minimum number of pre-analytic steps, to facilitate earlier intervention of patients suffering from

C. difficile-associated disease. Earlier intervention can also lead to more effective implementation of infection control measures, which can prevent further transmission to additional patients.

About the cobas® 4800 System
The cobas® 4800 System offers true walk-away automation of nucleic acid purification, PCR set-up and real-time PCR amplification and detection to help laboratories achieve maximum efficiency. The expanding system menu in the U.S. currently includes the cobas® MRSA/SA Test, cobas® CT/NG Test (Chlamydia trachomatis/Neisseria gonorrhoeae), cobas® HPV Test, cobas® BRAF V600 Mutation Test, cobas® EGFR Mutation Test and cobas® KRAS Mutation Test.

“With the addition of the cobas® Cdiff Test to the cobas® 4800 System menu, Roche is able to expand the tools available to assist clinicians in the management of healthcare associated infections,” said Paul Brown, head of Roche Molecular Diagnostics. “The cobas® Cdiff Test requires less sample handling and provides laboratories with a simplified workflow, when compared to other molecular methods. It also delivers a lower inhibition rate, which means fewer repeat samples and chances for error, enabling better patient care.”


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