Question 4-5: Were tests for molecular markers performed (e.g. PCR)?

Molecular markers for disease refer to specific genetic sequences which are believed to be associated with the recipient’s primary disease. Testing for these sequences is often performed using PCR based methods; however, lower sensitivity testing, including FISH, may also be used to detect molecular markers. Once a marker has been identified, these methods can be repeated to detect minimal residual disease (MRD) in the recipient’s blood, marrow, or tissue.

If testing for molecular markers was performed at the time of best response, report “yes” and indicate the sample collection date in question 5. If the exact date is not known, use the process described for reporting partial or unknown dates in General Instructions, General Guidelines for Completing Forms.

If no molecular marker testing was performed or it is unknown if testing was done, report “no” or “unknown” respectively and skip questions 5-13.

Question 6-13: Specify results

For each molecular marker in questions 6-12, report whether testing was “positive,” “negative,” or “not done.” If tests identified a molecular marker other than those listed in questions 6-11, report the result in question 12 and specify the marker in question 13.

If multiple “other molecular markers” were tested at the time of best response, report “see attachment” in question 13 and attach the final reports for any other markers which were tested. For further instructions on how to attach documents in FormsNet3SM, refer to the Training Guide. In this scenario, report “positive” in question 12 if any of the “other molecular markers” were detected.

Question 14-15: Was the disease status assessed via flow cytometry (minimum 4 color flow) (immunophenotyping)?

Flow cytometry (immunophenotyping) is a technique that can be performed on blood, bone marrow, or tissue preparations where cell surface markers can be detected on cellular material.

If flow cytometry (immunophenotyping) was performed at the time of best response, report “yes” and indicate the sample collection date in question 15. If the exact date is not known, use the process described for reporting partial or unknown dates in General Instructions, General Guidelines for Completing Forms.

If flow cytometry (immunophenotyping) was not performed, report “no” and skip questions 15-16.

Question 16: Was disease detected?

Indicate whether disease was detected by flow cytometry. If this is not clear from the laboratory report, consult with a physician and have them document whether evidence of disease is present.

Question 17: Was the disease status assessed via cytogenetic testing (karyotyping or FISH)?

Cytogenetic analysis is the study of chromosomes. Cytogenetic assessment involves testing blood or bone marrow for the presence of a known chromosomal abnormality which reflects the recipient’s disease. Testing methods you may see include conventional chromosome analysis (karyotyping) or fluorescence in situ hybridization (FISH). For more information about cytogenetic testing and terminology, see Appendix C.

Karyotyping is performed by culturing cells (growing cells under controlled conditions) until they reach the dividing phase. Techniques are then performed to visualize the chromosomes during cell division so that various bands and reconfigurations can be seen. Banding pattern differentiation and chromosomal reconfiguration demonstrate evidence of disease.

FISH is a sensitive technique that assesses a large number of cells. This technique uses special probes that recognize and bind to fragments of DNA commonly found in CLL. These probes are mixed with cells from the recipient’s blood. A fluorescent “tag” is then used to visualize the binding of the probe to the diseased cells. Additionally, the FISH probe panel should reflect the patient’s current disease; FISH may be used as surveillance for changes associated with post-therapy malignancy.

FISH testing for sex chromosomes after sex-mismatched allogeneic HCT should not be considered a disease assessment as the purpose is to determine donor chimerism.
If cytogenetic (karyotyping or FISH) studies were obtained at the time of best response, report “yes” and continue with question 18.

If cytogenetic studies were attempted at the time of best response, but there were not adequate cells (metaphases), report “no,” and skip questions 18-26.

If no cytogenetic studies were obtained at the time of best response, indicate “no” and skip questions 18-26.

Question 18-19: Was the disease status assessed via FISH?

If FISH studies were performed at the time of best response to HCT or cellular therapy, report “yes” and indicate the sample collection date in question 19. If the exact date is not known, use the process for reporting partial or unknown dates as described in the General Instructions, General Guidelines for Completing Forms.

If FISH studies were not performed, indicate “no” and skip questions 19-20. Examples of this include: no FISH study performed or FISH sample was inadequate.

Question 20: Was disease detected?

Indicate whether disease was detected by FISH. If the findings are unclear, consult with a physician and have them document whether evidence of disease is present.

Question 21-22: Was the disease status assessed via karyotyping?

If karyotyping (conventional cytogenetic) studies were performed at time of best response to HCT or cellular therapy, report “yes” and indicate the date sample collection date in question 22. If the exact date is not known, use the process for reporting partial or unknown dates as described in the General Instructions, General Guidelines for Completing Forms.

If karyotyping was not performed, report “no” and skip questions 22-23. Examples of this include: no karyotyping was performed or karyotyping culture failed.

Question 23: Was disease detected?

Indicate whether disease was detected by conventional cytogenetics (karyotyping). If the findings are unclear, consult with a physician and have them document whether evidence of disease is present.

Question 24-25: Was the disease status assessed by clinical/hematologic assessment?

Clinical and hematologic assessments are the least sensitive methods of establish a patient’s disease status. Examples include: pathologic evaluation (e.g., bone marrow biopsy), radiographic examination (e.g., X-ray, CT scan, MRI scan, PET scan), and laboratory assessment (e.g., CBC, peripheral blood smear), in addition to clinician evaluation and physical examination.

Indicate whether clinical and/or hematologic assessments were performed at the time the best response. If “yes,” report the date of assessment in question 25. The date reported should be that of the most-disease specific assessment performed at the time of best response (question 3). When determining the most disease-specific assessment, only consider studies which have previously or currently show evidence of disease. If all assessments are negative, report the date of the most sensitive test performed (e.g., report a bone marrow biopsy rather than a CBC) within the appropriate time period. If assessments are positive for disease, report the first assessment confirming the best response (question 1). If the exact date is not known, use the process for reporting partial or unknown dates as described in the General Instructions, General Guidelines for Completing Forms.

If no clinical and/or hematologic assessments were performed during this time period, report “no” and skip questions 25-26. This option should rarely be reported given the inclusion of physician assessments.

Question 26: Was disease detected?

Indicate whether disease was detected by clinical/hematologic assessment. If the findings are unclear, consult with a physician and have them document whether evidence of disease is present.

Last modified: Jun 30, 2017

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