October-November
Shelly has shown the pictures to colleagues at Exeter, but has urgent family issues to deal with. So I'll be waiting a while.

October
Have had a quick look at the reports and pictures with Shelly, and she intends to take them to her work colleagues for further views. Thence we will look at the slides.

September 19th
Spoke to Shelly and will call her when I get the piccies.

September 17th
Spoke to Laura and she said the loan of the slides is to avoid them lending me the microscope etc.

September 16th
Yesterday I paid to have pictures taken of the dodgy cells in my June tests.

The option will be to:

• Take more pictures of the slides: Must check my compact flash cards are viable.
• Take the slides to Exeter for further analysis.
• Do another cytology test. Have to chase up the staining procedure.

From : Roger Lovejoy
User-Agent: Thunderbird 2.0.0.14 (Windows/20080421)
To: plamkom@labcore.com
Subject: cytology and haematuria

Hi. Matthew

I just spoke to you from the UK about the ability to assess the origin of red blood cells by microscopic analysis. I had found this option via an associated website of yours: http://www.carepathonline.com/Topic.aspx?Nav=0&Rel=166

"If red blood cells are present in your urine sample, the pathologist will examine them closely with a high-powered microscope to observe their shape. The shape of the cells tells the pathologist whether they came from the kidney or from another part of your urinary tract."

As I mentioned the common options available to the punter, here is rather sketchy and it has taken me some months to get this far. Even the medical professionals seem to be bemused at my discoveries. (BTAStat, NMP22) and now this. The only view on cytology that I have extracted locally is being used to find cancer cells. My query is to know where the blood is coming from at this stage.

My queries are:

1. Is the analysis by an optical microscope or it some other higher tech. apparatus needed.
2. Can you provide me with any examples of how you discover the source, i.e. the differences in red blood cells. A description and/or images would be great. If it is done using an optical microscope I am thinking of buying one and doing regular checks myself.

Please let me know you get this so I can confirm I have the correct email address.

All the best

Roger Lovejoy

I haven't managed to assess this yet. I called Mandy Miller thinking she was working with the suppliers in Slough. I haven't received a reply, so today I have emailed nmp22@matritech.com. -:- Must call Mandy Miller of ‘Kyowa Hakko’ again (01753 566020)

16th May 10:07 Have just called Mandy wasn't available but someone else will look into it for me. Will call Germany Found 0049 761 478330. (http://www.matritech.de/) See if I can speak to Joe Hephler again.

Joe wasn't available but it was confirmed that the NMP22 uses the same goat - anti mouse derivative. I was told that no immunoassay can function without some animal derived antigen.

I have discovered that the BTA Stat test uses animal derivatives in its user-friendly monitor. I will be contacting them about this, but it looks like I won't be using it.

I had wondered a while ago how the monitor tested for a specific biological activity and although it may not use an animal derivative to test it does to produce a reference point.
 

The procedural Control (3) zone contains an immobilized goat anti-mouse IgG-specific antibody . . .
Bladderwatch BTAstat data.doc (1.1Mbytes)

8th May 16:25
Hav ejust emailed Nick Darker querying the above

1. Nuclear Matrix Protein 22 (NMP22) What is it? cancerbackup.org.uk

" NMP22 stands for nuclear matrix protein 22. Bladder cancer cells release this protein into urine. So NMP22 is usually found at higher levels in the urine of people with bladder cancer than in the urine of healthy people. However, not all samples of urine contain high levels of NMP22 even when there is cancer in the bladder. This means the test may give a negative result even if there is a cancer. This is called a false negative result. A false negative result is more likely for early bladder cancers and for bladder cancers that are slow growing (low grade). "

2. Comparison of NMP22 and Cytology. ingentia.com

" Results: Of the 46 recurrences detected by cystoscopy, the NMP22 test was positive in 39 cases and cytology in 19 cases. The sensitivity of the NMP22 test was 85%, which was significantly greater than that of cytology (41%). In particular, for low-risk tumors it was eight times more sensitive than cytology. The specificities of the NMP22 test and cytology were 77 and 96%, respectively. Combining the two tests increased overall sensitivity to 91%. However, 9% of the tumors were still not detected.

Conclusion: The NMP22 BladderChek test is an in vitro qualitative test that is easily available and cheap; it can be performed by a urologist in the office and results can be interpreted within 30 min. The NMP22 test is superior to cytology for all grades and stages in the detection of recurrence in patients with a history of superficial bladder cancer. Our study indicates that the NMP22 test can be used as a substitute for urine cytology. The NMP22 test cannot replace cystoscopy, but it can be used as an adjunct to cystoscopy in the surveillance protocol for patients with superficial bladder cancer. "

3. Screening and monitoring for bladder cancer: refining the use of NMP22.

" In the 135 patients with increased NMP22 values the 46 identified tumors were accompanied by 89 false-positive values yielding a specificity of 83.9% and a positive predictive value of 34.1%. These false-positive results were divided into 6 clinical categories. Exclusion of these categories improved the specificity and positive predictive value of NMP22 to 99.2% and 92.0%, respectively, yielding results similar to urinary cytology (99.8% and 94.1%).
Conclusion: Awareness and exclusion of the categories of false-positive results can increase the specificity and positive predictive value of NMP22, enhancing the clinical use of this urinary tumor marker. "

4. library.nhs.uk

" What were the findings?
Bladder cancer was diagnosed in 79 of the 1,331 patients. The NMP22 test had a sensitivity (the proportion of patients with the disease who tested positive) of 56% and a specificity (the proportion of patients without the disease who tested negative) of 86%. Cytology had a sensitivity of 16% and a specificity of 99%. The NMP22 test detected four cancers that were not seen during the first cystoscopy but were detected on subsequent examinations, including three that were muscle invasive and one carcinoma in situ.

What were the authors' conclusions?
The non-invasive point-of-care assay for elevated urinary NMP22 protein can increase the accuracy of cystoscopy, with test results available during the patient visit.

The results of this study also show that the NMP22 assay had significantly lower specificity (86%) than cytology (99%). This means that around 14% of people without bladder cancer were diagnosed as having bladder cancer on the NMP22 test, compared to 1% on the cytology test, which indicates that cytology could be more useful as a single test for ruling in a diagnosis of bladder cancer. "

From: clinchem.org

" Bladder cancer (BC)1 is the fourth most common cancer and cause of cancer-related deaths in men and the eighth in women; the 5-year survival rate of BC ranges between 30% and 90% (1). The main reasons for the poor survival despite the availability of effective treatment are a high frequency of recurrence (50–80% depending on the initial stage and grade) and lack of a sensitive method for early detection, particularly carcinoma in situ tumors. BC is usually associated with hematuria, dysuria, and frequent urination, but more specific screening tests for BC have not been available until recently.

Several BC tumor markers have been identified recently (2)(3)(4)(5). Among them, bladder tumor antigen (BTA) in urine is a sensitive marker and has been used as an adjunct to cystoscopy in the diagnosis and follow-up of patients with BC (6)(7)(8)(9)(10). BTA statTM and BTA TRAKTM (Alidex) are 2 commercially available assays for BTA in urine. Both assays use 2 monoclonal antibodies (mAbs) that bind to complement factor H (FH)-related material isolated from urine of BC patients (11). The BTA stat is a qualitative cartridge-form enzyme immunoassay, and the BTA TRAK is a quantitative sandwich-type immunoassay in 96-well format. In both assays, the mAb X52.1 is used as the capture antibody and X13.2 as the detection antibody. "

From: karger.co

" The BTA stat is a qualitative test which can be performed in a consultation setting. The BTA TRAK is a quantitative test that is performed in the laboratory. Consecutive patients highly suspicious of bladder cancer were included in this prospective blinded trial to assess the clinical performances of the two methods. Results: A total of 81 patients were tested using BTA stat and BTA TRAK before cystoscopy. A tumor was identified in 49 patients. "