JISCMail - ACB-CLIN-CHEM-GEN Archives

you could screen urines with the Freelite assay, although only immunofixation would indeed confirm that the free light chains are monoclonal and not ladder type.

I use the following approach:

It is well known that the usual dye binding methods for urine proteins may miss some BJP. However TCA is capable of precipitating ALL proteins and peptides. So we screen urines for proteins at a 30 mg/L cutoff with a modified TCA turbidimetric assay as follows:

1) prepare a standard by appropriately diluting to 30 mg/L a serum of known protein concentration.

2) sample & standard : 1 mL + 0.5 mL 15% TCA

3) sample blank & standard blank :1 mL +0.5 mL 0.1 N HCl

4) wait 10 min . room T

5) read against sample & standard blank at 405 nm

6) Fix or (Freelite) all samples that show an absorbance greater than the standard.

We have evaluated this approach ( unpublished results ) on over 100 urines with and without PBJ for wich the results of IF and Freelite assay were known. In no case the TCA screening missed a BJP.

----- Original Message -----

From: [log in to unmask] href="mailto:[log in to unmask]">Tilbrook Louise (RQ8) Mid Essex Hospital

To: [log in to unmask] href="mailto:[log in to unmask]">[log in to unmask]

Sent: Thursday, July 10, 2008 3:37 PM

Subject: Urine proteins and immunofixation

We are currently looking at rationalising our urine protein electrophoresis and immunofixation workload. In particular I'm concerned about the high number of fixes we do on specimens with very low amounts of urine protein and minimal staining on electrophoresis.

I haven't been able to find any guidelines or recent audits which address this issue - specifically how you adopt a pragmatic approach to detect as many BJPs as possible without investigating everything under the sun. I have found guidelines which suggest that BJP >0.03 ng/ml is regarded as clinically significant, others suggest >0.06.

If anyone has undergone this process recently I would be most grateful if they would be willing to share their experiences.

Principal Clinical Scientist

Mid Essex Hospitals NHS Trust

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