BD ProEx™ C Reagent

Frequently Asked Questions

BD ProEx™ C IHC (for the ICC FAQs, click here)

Questions		Answers

Test Request and Laboratory Testing

1. Can anyone in my lab perform the BD ProEx C IHC test?		IHC and ICC assays are categorized by CLIA as high-complexity tests. Test personnel must meet all criteria for labs performing high-complexity testing. Local and state regulations may also apply.

2. Does the patients doctor order this test?		No. BD ProEx C reagent in an IHC assay is ordered as part of the pathologist self-referral process. Pathologists order the test, not the patients physician. Pathologists often order ICC and IHC procedures in addition to other tests to make or confirm a diagnosis.

3. On what types of cases should I use BD ProEx C reagent?		BD ProEx C reagent when used with general use reagents in an IHC assay provides adjunctive information for histology diagnosis. Laboratories must validate the performance and clinical utility of BD ProEx C for their patient population and in so doing will determine the appropriate applications for the BD ProEx C test.

Specimen Handling

4. Can BD ProEx C reagent be used on frozen tissue specimens?		The current IHC protocol is only recommended for use with formalin-fixed paraffin-embedded tissue specimens.

5. Can BD ProEx C reagent be used on other types (non-cervical) of formalin-fixed paraffin-embedded tissue specimens?		Yes. Although we have not tested the performance of BD ProEx C reagent on other tissue specimens it should stain the nuclei of aberrant S-Phase cells in other tissue types.

Staining with ProEx C Reagent

6. How many slides can I stain with a vial of BD ProEx C reagent?		The number of slides that can be stained in an IHC procedure with a single vial of BD ProEx C Antibody is protocol dependent. Refer to the Package Insert 779-06555-00 for the recommended staining procedure with Ventanas iView reagents on the Benchmark XT and 779-06555-01 for the recommended staining procedure with Dakos Envision+ reagents on an open platform automated stainer.

7. What happens if the BD ProEx C reagent is frozen?		We have concluded that there is no effect on reagent performance when reagents are frozen, thawed, and tested immediately. However, we do not have data on long-term stability of the reagents when temperature conditions exceed the storage temperatures indicated on our labels and data sheets (refer to the Package Inserts 779-06555-00 and 779-06555-01).

8. What is the stability of the BD ProEx C antibody reagent after a bottle has been opened?		After opening the vial, the product is stable for 90 days when stored at the indicated temperature (2-8oC).

9. Can BD ProEx C reagent be used with the BD SureDetect™ reagents to stain formalin fixed paraffin embedded tissue specimens?		Yes, but we have not optimized or validated the BD SureDetect reagent protocol for tissue. The laboratory should validate the performance characteristics of the product prior to use.

10. Can I use other brands of visualization reagents and detection chemistry (other than Dakos Envision+ and Ventanas iView reagents) for BD ProEx C IHC?		BD ProEx C reagent was optimized for use with Dakos Envision+ and Ventanas iView detection reagents. If you want to use other visualization reagents then you are responsible for selecting the general use reagents, developing the test protocol, and validating performance characteristics.

11. Can I use the BD ProEx C reagent on other automated stainers?		Yes. BD ProEx C reagent is compatible for use on other open platform automated stainers.

Interpretation

12. Is a scoring guide or decision tree available when looking a BD ProEx C IHC slides?		No, unlike the BD ProEx C cytology package insert there is no basic scoring tree available for BD ProEx C IHC. In tissue, the pathologist is examining the staining of the cells in situ and scoring algorithms are not necessary.

13. What should I do if a normal cell stains positive with the BD ProEx C IHC method?		TOP2A and MCM2 are expressed in some proliferating cells (particularly basal cells) and therefore it is possible to see some staining in tissue. However, the anatomic pathologist is able to distinguish between normal cell staining patterns and atypical cell staining patterns in tissue.

14. Does BD ProEx C reagent stain tissue just like p16?		No, the staining patterns are different. BD ProEx C is an antibody reagent containing antibodies to MCM2 and TOP2A proteins. The p16 reagent is an antibody specific to p16. The BD ProEx C IHC produces a nuclear staining pattern while the p16 antibody stains both the nucleus and cytoplasm. Abstract 928 from the 2005 USCAP conference in the Clinical Reference Folder (980-06163-00) describes the research studies correlating p16, MCM2, and Cyclin E as molecular markers in IHC and ICC formats.

Reimbursement

15. Is the BD ProEx C IHC method reimbursable?		Yes. Immunocytochemistry and Immunohistochemistry procedures with manual interpretation are reimbursable under the 88342 CPT code.

BD ProEx™ C ICC (for the IHC FAQs, click here)

Test Request and Laboratory Testing

1. Can anyone in my lab perform the BD ProEx™ C ICC test?		ICC and IHC assays are categorized by CLIA as high-complexity tests. Test personnel must meet all criteria for labs performing high-complexity testing. Local and state regulations may also apply.
2. Does the patient’s doctor have to order this test?		No. BD ProEx™ C reagent in an ICC assay is ordered as part of the pathologist self-referral process. Pathologists order the test, not the patient’s physician. Pathologists often order ICC and IHC procedures in addition to other tests to make or confirm a diagnosis.
3. What types of cases should I reflex to BD ProEx™ C reagent?		BD ProEx™ C reagent when used with general use reagents in an ICC assay provides adjunctive information for cytology diagnosis. Laboratories must validate the performance and clinical utility of BD ProEx™ C reagent for their patient population and in so doing will determine the appropriate applications for the BD ProEx™ C reagent test. (Reference: Updates in Molecular Diagnostics Folder, 980-0163-00, Freeman et al. Clinical Cancer Research, 1999).

Specimen Handling

4. Can I prepare and store slides from cytology specimens in advance of staining with BD ProEx™ C reagent?		No. Cytology specimens may be stored in the refrigerator and processed immediately before staining, but the slides cannot be air dried and stored for future staining.
5. How long can I keep my cytology samples before staining with BD ProEx™ C reagent?		Refer to the manufacturer’s instructions for the specimen collection and handling method. The laboratory should validate the performance characteristics of the product prior to use.

Staining with ProEx™ C reagent

6. What is the stability of the BD ProEx™ C antibody reagent after a bottle has been opened?		After opening the vial, the product is stable for 90 days when stored at the indicated temperature (2-8C).
7. How many slides can I stain with a vial of BD ProEx™ C reagent?		If you use 200 mL of BD ProEx™ C reagent on each slide, you will be able to stain 75 slides with a single vial. Refer to the Protocol section on the BD ProEx™ C Package Insert (779-11000-20).
8. What happens if the BD ProEx™ C antibody reagent is frozen?		We have concluded that there is no effect on reagent performance when reagents are frozen, thawed, and tested immediately. However, we do not have data on long-term stability of the reagents when temperature conditions exceed the storage temperatures indicated on our labels and data sheets (Reference BD ProEx™ C Package Insert; 779-11000-20; carton and vial labels).
9. Can I use my own brands of visualization reagents and detection chemistry with BD ProEx™ C reagent?		BD ProEx™ C reagent was optimized for use with the BD SureDetect™ general use reagents. If you want to use other visualization reagents then you are responsible for selecting the general use reagents, developing the test protocol, and validating performance characteristics.

Interpretation

10. Is a scoring guide or decision tree available when looking at BD ProEx™ C ICC slides?		Yes, there is a basic scoring tree available in the BD ProEx™ C Package Insert (779-11000-20). However, it is the responsibility of the laboratory to develop and validate any scoring guides or decision trees for assay interpretation. BD Diagnostics - TriPath has a Slide Atlas (785-06591-00) that includes examples of cytology slides that have stained positive or negative for aberrant S-Phase induction. These examples may be useful in the development of your own scoring guide.
11. What should I do if a normal cell stains positive with the BD ProEx™ C ICC method?		TOP2A and MCM2 are expressed in some proliferating cells and therefore it is possible to see some staining in morphologically normal appearing cells. However, the biomarkers are typically over-expressed in atypical cells. Therefore normal appearing cells that are positive for the markers should be reported as negative.
12. IWill the cytoplasm ever stain with BD ProEx™ C ICC methods?		The expected staining pattern for BD ProEx™ C reagent in an ICC format is nuclear. Cytoplasmic staining may be the result of a problem with the specimen or the ICC protocol.
13. What terminology should be used on the cytology report to report the BD ProEx™ C ICC result?		BD ProEx™ C reagent results are either positive or negative for the detection of aberrant S-Phase induction. The annotation included in the report is laboratory specific.
14. Is BD ProEx™ C reagent just like p16?		No, the staining patterns are different. BD ProEx™ C is a reagent containing antibodies to MCM2 and TOP2A proteins. The p16 reagent is an antibody specific to p16. BD ProEx™ C reagent produces a nuclear staining pattern while the p16 antibody stains both the nucleus and cytoplasm. The 2005 USCAP conference in the Clinical Reference Folder (980-06163-00) describes the research studies correlating p16, MCM2, and Cyclin E as molecular markers in IHC and ICC formats.

Reimbursement

15. Is the BD ProEx™ C ICC method reimbursable?		Yes. Immunocytochemistry and Immunohistochemistry procedures with manual interpretation are reimbursable under the 88342 CPT code.