Impact of 2013 American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) guidelines on HER2 fluorescent in situ hybridization (FISH) testing in breast cancers: Experience from a national reference laboratory.

e23188 Background: Accurate HER2 testing is expected to identify patients who will benefit from HER2 directed therapy (HDT) minimizing false positive and negatives. Like many biologic processes, HER2 protein expression and gene copy numbers are a continuum, however these tests are expected to be reported dichotomously. In 2013, ASCO/CAP revised the guidelines in an attempt to decrease false negative results and reverted to FDA approved ratio of ≥ 2.0 for HER2 positivity. We reviewed HER2 FISH testing results after the implementation of 2013 ASCO/CAP guidelines to determine the effects on HER2 reporting from our national reference laboratory. Methods: HER2 FISH testing performed between 5/2015-4/2016 at ARUP Labs following current 2013 ASCO/CAP guidelines was included. HER2 to control probe ratios, mean HER2 and control probe copy numbers were used to reassign HER2 status using 2007 ASCO/CAP, and FDA guidelines for each case. Results: HER2 FISH results were available in 2,017 cases. 342 (17.0%) cases were amplified, 301 (14.9%) were equivocal, and 1374 (68.1%) were non-amplified. After additional testing with alternate probe, 93 (31.2%) of the equivocal cases were reclassified as amplified increasing amplified cases to 21.6%. All of the equivocal cases which were reinterpreted as amplified with alternate probe showed low level amplification (range: 2.0-3.6; mean: 2.3). HER2 positivity rates following 2013 ASCO/CAP guidelines, both at initial testing and after additional testing to resolve equivocal results were significantly higher compared to 2007 ASCO/CAP guidelines and FDA criteria. Conclusions: 2013 ASCO/CAP guidelines lead to higher number of HER2 FISH positive and equivocal cases. In a reference laboratory setting where alternative control probe was used to resolves equivocal FISH cases, 31.2% of patients with initial equivocal results become HER2 positive. However, it is not known if these patients benefit from targeted therapies as they would not be included in the original adjuvant or metastatic trials.

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Evaluation of optimal urine screening and confirmation cut-off values for opiates, at a national reference laboratory

Turkish Journal of Biochemistry ◽

10.1515/tjb-2020-0614 ◽

2021 ◽

Vol 0 (0) ◽

Author(s):

Çiğdem Karakükcü ◽

Mehmet Zahid Çıracı ◽

Derya Kocer ◽

Mine Yüce Faydalı ◽

Muhittin Abdulkadir Serdar

Keyword(s):

False Positive ◽

False Positive Rate ◽

False Negative ◽

Urine Samples ◽

Reference Laboratory ◽

National Reference Laboratory ◽

Urine Screening ◽

National Reference ◽

Positive Rate ◽

Sensitivity Specificity

Abstract Objectives To obtain optimal immunoassay screening and LC-MS/MS confirmation cut-offs for opiate group tests to reduce false positive (FP) and false negative (FN) rates. Methods A total of 126 urine samples, −50 opiate screening negative, 76 positive according to the threshold of 300 ng/mL by CEDIA method – were confirmed by a full-validated in-house LC-MS/MS method. Sensitivity, specificity, FP, and FN rates were determined at cut-off concentrations of both 300 and 2,000 ng/mL for morphine and codeine, and 10 ng/mL for heroin metabolite 6-mono-acetyl-morphine (6-MAM). Results All CEDIA opiate negative urine samples were negative for morphine, codeine and 6-MAM. Although sensitivity was 100% for each cut-off; specificity was 54.9% at CEDIA cut-off 300 ng/mL vs. LC-MS/MS cut-off 300 ng/mL and, 75% at CEDIA cut-off 2,000 ng/mL vs. LC-MS/MS cut-off 2,000 ng/mL. False positive rate was highest (45.1%) at CEDIA cut-off 300 ng/mL. At CEDIA cut-off 2,000 ng/mL vs. LC-MS/MS cut-off 300 ng/mL, specificity increased to 82.4% and FP rate decreased to 17.6%. All 6-MAM positive samples had CEDIA concentration ≥2,000 ng/mL. Conclusions 2,000 ng/mL for screening and 300 ng/mL for confirmation cut-offs are the most efficient thresholds for the lowest rate of FP opiate results.

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American Society of Clinical Oncology/College of American Pathologists 2018 Focused Update of Breast Cancer HER2 FISH Testing GuidelinesResults From a National Reference Laboratory

American Journal of Clinical Pathology ◽

10.1093/ajcp/aqz061 ◽

2019 ◽

Vol 152 (4) ◽

pp. 479-485 ◽

Cited By ~ 4

Author(s):

Leo Lin ◽

Deepika Sirohi ◽

Joshua F Coleman ◽

H Evin Gulbahce

Keyword(s):

Breast Cancer ◽

Metastatic Breast ◽

Reference Laboratory ◽

National Reference Laboratory ◽

Breast Cancers ◽

National Reference ◽

American Society ◽

New Guidelines ◽

Her2 Positivity

Abstract Objectives To review impact of the ASCO/CAP 2018 update on HER2 testing. Methods HER2 fluorescence in situ hybridization (FISH) test requests from primary and metastatic breast cancers between August 2018 and January 2019 were included. FISH results requiring a changed algorithm under the new guidelines (groups 2, 3, and 4) were identified and HER2:CEN17 ratios, average HER2, CEN17 signals/cell, and HER2 immunohistochemistry (IHC) results were recorded. Results Of the HER2 FISH cases 176/812(21.7%) fell within groups 2, 3, or 4; 0/12, 1/12, and 2/152 cases were positive (3+) by IHC, and 1/12, 2/12, and 6/152 cases were positive after targeted scoring from groups 2, 3, and 4, respectively. Following 2018 updates, 8.3%, 25%, and 5.3% of the groups 2, 3, and 4 were positive, respectively. Conclusions Groups 2, 3, and 4 constituted over 20% of HER2 FISH tests in a reference laboratory. The 2018 ASCO/CAP update significantly decreased the HER2 positivity rate.

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Reassignment of HER2 status for subgroups of breast cancer according to the 2018 updated American Society of Clinical Oncology and College of American Pathologists guidelines: The impact of combined immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) reflex testing in a large national reference laboratory.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.3144 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. 3144-3144

Author(s):

Katherine Geiersbach ◽

Reid G. Meyer ◽

Sara M. Kloft-Nelson ◽

Darlene L. Knutson ◽

Ryan A. Knudson ◽

...

Keyword(s):

Breast Cancer ◽

Her2 Status ◽

Reference Laboratory ◽

National Reference Laboratory ◽

National Reference ◽

Group 4 ◽

Reflex Testing ◽

Group 3 ◽

Group 2 ◽

The Impact

3144 Background: Updated ASCO/CAP Guidelines for HER2 testing in breast cancer have been most impactful on the resolution of certain challenging groups of FISH results. We review the change in assignment of HER2 status in a large series of breast cancers referred to a large national reference laboratory for FISH testing since the introduction of the 2018 updated guidelines. Methods: Patient samples submitted to the Mayo Clinic Cytogenetics Laboratory (N = 2208) were analyzed by FISH. Samples with Group 2, Group 3, or Group 4 FISH results were reflexed to immunohistochemistry (IHC) in our central laboratory; FISH slides for those cases with equivocal 2+ IHC results were re-scored in the regions of invasive cancer showing more intense membranous staining. A subset of 202 samples with Group 4 FISH results were also reflexed to the previously employed reflex FISH assay (HER2/D17S122), and these were also re-analyzed according to the new reflex IHC/FISH process. Results: 382 of 2208 breast cancer samples tested (17.3%) had FISH results categorized as Group 2 (N = 17, 0.8%), Group 3 (N = 34, 1.5%), or Group 4 (N = 331, 15%) and required reflex IHC testing, and of those, 75% were 2+ equivocal and required targeted re-analysis of the FISH slide according to the 2018 updated guidelines. Re-analysis of the FISH slide resulted in switching between Groups 1-5 in 19.4% of cases, but HER2 status was changed by FISH re-scoring in only 7.7% of cases re-scored (1.0% of all samples), generally due to only minor shifts in HER2 copy number and HER2/control ratios between the initial and IHC-guided reflex FISH scores. In the subset of 202 cases tested by both reflex methods, the previously employed HER2/D17S122 reflex probe set was positive in 123 cases (60.9%), whereas reflex IHC/FISH was positive in only 10 cases (7.9%). Including positive reflex IHC (0.4%) and positive reflex FISH results (2.1%), the overall assignment of positive HER2 status on our series of 2208 cases was 11.5%. Conclusions: Overall rates of HER2 positive FISH results have declined under the most recent ASCO/CAP guideline update as a consequence of new recommendations for reflex testing for Groups 2-4. This change is largely due to reassignment of Group 2 and Group 4 results as negative in the absence of positive IHC.

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RAI1 Alternate Probe Identifies Additional Breast Cancer Cases as Amplified Following Equivocal HER2 Fluorescence In Situ Hybridization Testing: Experience From a National Reference Laboratory

Archives of Pathology & Laboratory Medicine ◽

10.5858/arpa.2016-0201-oa ◽

2016 ◽

Vol 141 (2) ◽

pp. 274-278 ◽

Cited By ~ 10

Author(s):

Ling Hui ◽

Katherine B. Geiersbach ◽

Erinn Downs-Kelly ◽

H. Evin Gulbahce

Keyword(s):

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

False Negative ◽

Metastatic Breast ◽

Her2 Status ◽

Reference Laboratory ◽

Breast Cancers ◽

Negative Results ◽

Control Probe

Context.—In 2013 the American Society of Clinical Oncology and College of American Pathologists updated the HER2 guidelines and changed the equivocal category for HER2 in situ hybridization testing to an average HER2 copy number of 4.0 to 5.9 with a HER2:CEP17 ratio of less than 2.0 and proposed retesting, with an option of using another control probe to avoid false-negative results. RAI1, located at band position 17p11.2, is a popular alternate probe locus for retesting equivocal changes. Objective.—To review experience with the RAI1 alternate probe in HER2 fluorescence in situ hybridization equivocal breast cancers. Design.—Primary and metastatic breast cancers with equivocal HER2 fluorescence in situ hybridization, retested with an alternate (RAI1) probe, were identified. HER2, RAI1, and CEP17 copy numbers, HER2 to control probe ratios, and genetic heterogeneity were recorded. Hematoxylin-eosin–stained slides were reviewed for type and grade of cancer. Results.—Of 876 cases tested with CEP17 as the reference probe, 97 (11.1%) had equivocal HER2 fluorescence in situ hybridization results. Additional testing with the RAI1 probe classified 39.2% cases (38 of 97) as amplified with a HER2:RAI1 ratio ranging from 2.0 to 3.2 (mean, 2.37); 3.1% (3 of 97) were still unclassifiable because of a deletion of RAI1. Conclusions.—RAI1 identified close to 40% of original HER2 fluorescence in situ hybridization equivocal cases as amplified, making these patients eligible for targeted therapies. It is not known whether guidelines for US Food and Drug Administration–approved probes can be extrapolated to alternate probes when an alternate control probe shows losses or gains. Because of the lack of guidelines for reporting HER2 status with alternate probes, laboratories face challenges in interpreting results.

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A survey of European National Reference Laboratory capacity for pertussis PCR and serology diagnostics, 2015

10.26226/morressier.57fe118dd462b8028d890950 ◽

2016 ◽

Author(s):

Norman Fry

Keyword(s):

Reference Laboratory ◽

National Reference Laboratory ◽

National Reference ◽

Laboratory Capacity

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Laboratory capacity of Greek hospitals for diagnosis of salmonellosis and surveillance systems’ performance in the years of economic crisis, 2010–2016

Epidemiology and Infection ◽

10.1017/s0950268818002686 ◽

2018 ◽

Vol 147 ◽

Author(s):

K. Mellou ◽

E. Saranti-Papasaranti ◽

G. Mandilara ◽

T. Georgakopoulou

Keyword(s):

Public Hospitals ◽

Geographical Region ◽

Reference Laboratory ◽

Laboratory System ◽

Surveillance Systems ◽

National Reference Laboratory ◽

Notification System ◽

National Reference ◽

Laboratory Capacity ◽

Mandatory Notification

AbstractAusterity might have affected the capacity of public hospitals in Greece to diagnose salmonellosis (laboratory capacity) over the period 2010–2016, as well as the performance of the existing surveillance systems. The scope of this paper is to present data on laboratory capacity over these years, as well as the results of a two-source capture-recapture study (data from Mandatory Notification System and National Reference Laboratory System for Salmonella). The main findings were that: (a) laboratory capacity was high and steady besides the financial crisis, (b) the estimated number of laboratory-confirmed cases (n = 6017, 95% CI 5892–6142) resulted in an incidence rate (7.9 cases/100 000 population) almost twice than that reported by the two systems Mandatory Notification System (MNS); 4.1 and National Reference Laboratory System (NRLS); 4.5 cases/100 000 population, (c) underreporting was high for both systems (MNS; 47.5% and NRLS; 42.8%) and (d) differences by geographical region, size and type of hospital were identified. We suggest that (a) specific interventions are needed to increase completeness of the systems by type of hospital and geographical region, (b) record linkage can help in estimating the disease burden in a more valid way than each system separately and (c) a common electronic database in order to feed one system to the other could significantly increase completeness of both systems.

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