Drug Administration-approved Test Research Articles

Self-Collection for HPV Testing: Potential Issues Related to Performance Measures and Quality Improvement Among Federally Qualified Health Centers.

Most incident cases of cervical cancer in the United States are attributable to inadequate screening. Federally qualified health centers (FQHCs) serve a large proportion of women who are low-income, have no insurance, and are underserved-risk factors for insufficient cervical cancer screening. FQHCs must maintain quality measures to preserve their accreditation, address financial reimbursements, and provide quality care. Implementation of human papillomavirus (HPV) self-collection can improve cervical cancer screening coverage within FQHCs. To understand perspectives from clinical personnel on current cervical cancer screening rates at FQHCs in North Carolina and the impact of implementing HPV self-collection among underscreened patients on screening rates and performance measures. The study used focus groups and key informant interviews. Coding-based thematic analysis was applied to both focus group and interview transcripts. Emergent themes regarding perspectives on self-collection implementation were mapped onto Consolidated Framework for Implementation Research (CFIR) constructs to identify future barriers and facilitators to implementation. Two FQHCs in North Carolina and a cloud-based videoconferencing platform. Six FQHCs in North Carolina; 45 clinical and administrative staff from the 6 FQHCs; 1 chief executive officer (n = 6), 1 senior-level administrator (n = 6), 1 chief medical officer (n = 6), and 1 clinical data manager (n = 6) from each FQHC. Achievement of clinical perspectives. Societal-, practice-, and patient-level factors currently contribute to subpar cervical cancer screening rates. HPV self-collection was expected to improve screening uptake among underscreened women at FQHCs, and thus quality and performance measures, by offering an alternative screening approach for in-clinic or at-home use. Implementation barriers include financial uncertainties and HPV self-collection not yet a Food and Drug Administration-approved test. HPV self-collection has potential to improve cervical cancer screening quality and performance measures of FQHCs. For a successful implementation, multilevel factors that are currently affecting low screening uptake need to be addressed. Furthermore, the financial implications of implementation and approval of HPV self-collection as a test for cervical cancer screening quality measures need to be resolved.

Olutasidenib: First Approval.

Olutasidenib (REZLIDHIATM), an isocitrate dehydrogenase-1 (IDH1) inhibitor, is being developed by Rigel Pharmaceuticals for the treatment of relapsed or refractory (R/R) acute myeloid leukaemia (AML). Olutasidenib was recently approved in the USA for the treatment of adults with R/R AML with a susceptible IDH1 mutation as detected by a US Food and Drug Administration-approved test. This article summarizes the milestones in the development of olutasidenib leading to this first approval for R/R AML.

KRAS-targeted therapy in the treatment of non-small cell lung cancer.

KRAS mutations are one of the most common driver mutations in non-small cell lung cancer. Though previously believed to be an undruggable target, recent advances in therapeutics have seen new targeted agents against KRAS mutations. The objective of this article is to review currently available and upcoming KRAS-targeted treatments. Currently available trials examining KRAS-targeted therapy in non-small cell lung cancer were examined by searching for the keyword "KRAS inhibitors." The pivotal trials for sotorasib and adagrasib were reviewed for this article. Mutated KRAS can be challenging to target for a variety of reasons. In 2021, the US Food and Drug Administration approved sotorasib for the treatment of adult patients with locally advanced or metastatic non-small cell lung cancer with KRAS G12C mutation as determined by a Food and Drug Administration-approved test, who have received at least one prior systemic therapy. A multicenter, single-group, open-label, phase 2 trial was able to demonstrate that sotorasib was able to demonstrate objective response, progression-free survival, and overall survival in this patient population. A phase 3 trial comparing sotorasib to docetaxel in the subsequent-line treatment of KRAS G12C non-small cell lung cancer is currently ongoing. There are other KRAS-targeted agents currently under study, including adagrasib, with growing interest in targeting KRAS downstream pathways. Further trials need to be conducted in order to identify other targeted agents for KRAS and the appropriate place in therapy among currently approved treatments for non-small cell lung cancer.

Validation and Characterization of FGFR2 Rearrangements in Cholangiocarcinoma with Comprehensive Genomic Profiling

Cholangiocarcinoma (CCA) is a heterogeneous biliary tract cancer with a poor prognosis. Approximately 30% to 50% of patients harbor actionable alterations, including FGFR2 rearrangements. Pemigatinib, a potent, selective fibroblast growth factor receptor (FGFR) FGFR1-3 inhibitor, is approved for previously treated, unresectable, locally advanced or metastatic CCA harboring FGFR2 fusions/rearrangements, as detected by a US Food and Drug Administration-approved test. The next-generation sequencing (NGS)-based FoundationOneCDx (F1CDx) was US Food and Drug Administration approved for detecting FGFR2 fusions or rearrangements. The precision and reproducibility of F1CDx in detecting FGFR2 rearrangements in CCA were examined. Analytical concordance between F1CDx and an externally validated RNA-based NGS (evNGS) test was performed. Identification of FGFR2 rearrangements in the screening population from the pivotal FIGHT-202 study (NCT02924376) was compared with F1CDx. The reproducibility and repeatability of F1CDx were 90% to 100%. Adjusted positive, negative, and overall percentage agreements were 87.1%, 99.6%, and 98.3%, respectively, between F1CDx and evNGS. Compared with evNGS, F1CDx had a positive predictive value of 96.2% and a negative predictive value of 98.5%. The positive percentage agreement, negative percentage agreement, overall percentage agreement, positive predictive value, and negative predictive value were 100% for F1CDx versus the FIbroblast Growth factor receptor inhibitor in oncology and Hematology Trial-202 (FIGHT-202) clinical trial assay. Of 6802 CCA samples interrogated, 9.2% had FGFR2 rearrangements. Cell lines expressing diverse FGFR2 fusions were sensitive to pemigatinib. F1CDx demonstrated sensitivity, reproducibility, and high concordance with clinical utility in identifying patients with FGFR2 rearrangements who may benefit from pemigatinib treatment.

Cutaneous leishmaniasis in Bikaner, India: Clinicoepidemiological profile; parasite identification using conventional, molecular methods and CL Detect™ rapid test, a new Food and Drug Administration-approved test.

Background and Objective:Cutaneous Leishmaniasis (CL) is a dermal manifestation caused by various species of Leishmania. This is the most common Leishmanial syndrome seen worldwide and is emerging as and threatens to become an uncontrollable disease. The present study was planned to understand the current epidemiology of CL in the conventionally endemic area of Bikaner, Rajasthan. Species characterization was also done.Material and Methods:The diagnostic modalities used in the study were microscopy, histopathologic examination, antigen testing and confirmation and species characterization by PCR. Field evaluation of the new antigen detection kit approved by FDA in 2016 CL Detect™ IC-RDT was done and its results compared with other available diagnostic tests.Results:A total of 14 cases with 25 skin lesions presented to the Skin OPD during the six month study period (January to June 2018). Out of these, 5 were males and 9 were females, average age being 32 yrs. Students and housewives of poor socio economic group were the most frequently affected groups. Lesions were found on the exposed areas of the body, mainly on the upper limbs and facial region. Majority of them were ulcerated plaques. Out of 14 cases under study, antigen test confirmed 7 positives; thereby yielding a positive rate of 50% against 71.4% for microscopy, 72.7% for PCR and a mere 38.5% for histopathology. Species characterization revealed Leishmania tropica as the causative organism.Interpretation and Conclusion:The study indicates that the number of cases have declined substantially over the past decade in Bikaner. Despite being the oldest technique used, microscopy still is a good option for diagnosis. Combined with PCR, diagnostic accuracy and hence utility is increased multifold. In its current form, IC-RDT may not be very useful.

Clinical Evaluation of the Cartridge-Based GeneXpert Human Papillomavirus Assay in Women Referred for Colposcopy

High-risk human papillomavirus (hrHPV) testing is now being introduced as a potential primary screening test for improved detection of cervical precancer and cancer. Current U.S. Food and Drug Administration-approved tests are batch tests that take several hours to complete. A rapid, non-batch test might permit point-of-care (POC) testing, which can facilitate same-day screen and management strategies. For a non-batch, random-access platform (GeneXpert; Cepheid, Sunnyvale, CA), a prototype hrHPV assay (Xpert) has been developed where testing for 14 hrHPV types can be completed in 1 h. In the first clinical evaluation, Xpert was compared to two validated hrHPV tests, the cobas HPV test (cobas, Roche Molecular Systems) and Hybrid Capture 2 (hc2, Qiagen), and to histologic outcomes using specimens from colposcopy referral populations at 7 clinical sites in the United States (n = 697). The sensitivity of Xpert for cervical intraepithelial neoplasia grade 2 or more severe diagnoses (CIN2+) (n = 141) was equal to that of cobas (90.8% versus 90.8%, P = 1) and greater than that of hc2 (90.8% versus 81.6%, P = 0.004). Xpert was more specific than cobas (42.6% versus 39.6%, P = 0.02) and less specific than hc2 (42.6% versus 47.7%, P < 0.001). Similar results were observed for cervical intraepithelial neoplasia grade 3 or higher (CIN3+) (n = 91). HPV16 detection by Xpert identified 41.8% of the CIN2+ specimens with a positive predictive value (PPV) of 54.6%. By comparison, HPV16 detection by cobas identified 42.6% of the CIN2+ specimens with a PPV of 55.0%. hrHPV detection by the Xpert demonstrated excellent clinical performance for identifying women with CIN2+ and CIN3+ that was comparable to that of currently available clinically validated tests.

Cost Effectiveness of Crizotinib for Anaplastic Lymphoma Kinase–Positive, Non–Small-Cell Lung Cancer: Who Is Going to Blink at the Cost?

In 2014, lung cancer is the poster child for personalized medicine, and identifying targetable oncogenic drivers is now considered standard of care in stage IV disease. The anaplastic lymphoma kinase (ALK) gene has emerged as an important oncogenic driver in a small population of patients with adenocarcinoma. Crizotinib has received accelerated US Food and Drug Administration approval when used in conjunction with its companion diagnostic test to identify patients with the EML4-ALK gene rearrangement. No one who treats patients with lung cancer questions the exciting results obtained with crizotinib, or the need for a highly specific test. A recent phase III study compared crizotinib with standard chemotherapy in patients with locally advanced or metastatic ALK-positive lung cancer who had received one previous platinum-based regimen. Everything favored crizotinib: median progression-free survival (PFS), 7.7 versus 3.0 months; response rate, 65% versus 20%; and symptoms and quality of life were also substantially better. Overall survival was not improved, but 64% of the group receiving chemotherapy crossed over to crizotinib. Two main questions remain unanswered: Can we afford to screen everyone with lung cancer, given that only 3% to 5% of the population will be ALK positive, and can we afford to pay for crizotinib? In the article prompting this editorial, Djalalov et al present their results of a cost-effectiveness analysis of both EML4-ALK fusion testing and first-line crizotinib using a Markov model in newly diagnosed, previously untreated patients with stage IV non–small-cell lung cancer (NSCLC) and nonsquamous histology. The authors compared strategies for treatment: crizotinib for patients who test ALK positive versus standard chemotherapy for those who test negative. The incremental cost-effectiveness ratio (ICER) or change in costs divided by the change in benefits is commonly used in health economics to provide a practical approach to decision making regarding health interventions. The costs are described in monetary units, whereas the benefits or effects on health status are measured in quality-adjusted life years (QALYs) gained or lost. Using a relative efficacy estimate in the first-line setting that was derived from the second-line phase III trial, the author’s projected that first-line crizotinib gives 0.64 years (7.7 months) extra survival, or 0.38 QALYs, at an additional cost of $95,000 compared with standard of care, leading to ICERs of $148,000 per additional life year, or $250,000/ QALY gained. The authors concluded that EML4-ALK fusion testing and crizotinib treatment for patients positive for ALK is not costeffective (from a Canadian perspective) because of the low biomarker frequency in the population—and the high drug costs. Are their conclusions valid, what do they mean, and is crizotinib affordable? The model seems valid. The authors simulated the clinical trial of what happens if patients positive for ALK start with crizotinib while we await the results of the ongoing first-line phase III PROFILE 1014 trial (A Clinical Trial Testing the Efficacy of Crizotinib Versus Standard Chemotherapy Pemetrexed Plus Cisplatin or Carboplatin in Patients With ALK Positive Nonsquamous Cancer of the Lung). In this analysis, all patients receive crizotinib (or do not, if ALK negative), then cisplatin plus gemcitabine, then second-line pemetrexed, then thirdline erlotinib, regardless of epidermal growth factor receptor status. The authors transform the added 0.64 years of life to 0.38 qualityadjusted years by assuming that the quality of life is only 3% better for patients receiving crizotinib compared with regular chemotherapy. This seems ungenerous, given the rapid and often dramatic symptom improvement seen in practice, which should translate into greater quality time while receiving the drug. Their test strategy for ALK assessment is efficient—a critically important aspect, given the need to screen 100 patients to find approximately three who test positive. The only US Food and Drug Administration–approved test is the Vysis LSI ALK Break Apart FISH Probe Kit (Abbott Molecular, Des Plaines, IL), with near 100% accuracy but at a cost of more than $250/test. The correlation of results of ALK immunohistochemistry (IHC) and ALK fluorescent in situ hybridization (FISH) are excellent, but IHC has not been validated as a predictive marker for response to an ALK inhibitor in a large patient cohort. The US guidelines state that ALK IHC, if carefully validated, may be considered for screening patients with lung adenocarcinoma. The authors used ALK IHC (approximately $40) with 95% sensitivity as their baseline, with subsequent verification with ALK FISH if needed. Djalalov et al indicate that EML4-ALK fusion testing with crizotinib treatment for patients positive for ALK “is not cost-effective in the setting of high drug costs and a low biomarker JOURNAL OF CLINICAL ONCOLOGY E D I T O R I A L VOLUME 32 NUMBER 10 APRIL 1 2014

Results of lookback for Chagas disease since the inception of donor screening at New York Blood Center

Chagas disease is a parasitic infection by Trypanosoma cruzi, typically transmitted via infected triatomine bug fecal contamination of bite sites. Other routes of infection include congenital, oral, organ transplantation, and blood product transmission. From 2007 until 2011, New York Blood Center screened donations for the presence of T. cruzi antibodies using a Food and Drug Administration-approved test. Confirmatory testing was performed and recipients of units donated by confirmed-positive donors were investigated via lookback. A total of 204 donors were T. cruzi antibody positive representing 0.019% of all donors during this time period (1,066,516 unique donors screened). Of the enzyme-linked immunosorbent assay-reactive donors, 77 were confirmed positive by radioimmunoprecipitation assay (0.007%). At least 154 units from 29 of the confirmed-positive donors had been transfused to 141 recipients. At the time of lookback, 48 of the 141 recipients were alive and seven underwent T. cruzi screening. Two recipients were found to be immunofluorescence assay (IFA) positive. Both IFA-positive recipients received a leukoreduced apheresis platelet unit (two separate donations) from the same confirmed positive donor, a 72-year-old immigrant from Argentina. Lookback analysis was able to identify the first two cases of probable transfusion-transmitted T. cruzi infection since implementation of the national screening program, which increases the total number of reported cases in the United States to 8.

Verification of Performance Specifications of a Molecular Test: Cystic Fibrosis Carrier Testing Using the Luminex Liquid Bead Array

The number of clinical laboratories introducing various molecular tests to their existing test menu is continuously increasing. Prior to offering a US Food and Drug Administration-approved test, it is necessary that performance characteristics of the test, as claimed by the company, are verified before the assay is implemented in a clinical laboratory. To provide an example of the verification of a specific qualitative in vitro diagnostic test: cystic fibrosis carrier testing using the Luminex liquid bead array (Luminex Molecular Diagnostics, Inc, Toronto, Ontario). The approach used by an individual laboratory for verification of a US Food and Drug Administration-approved assay is described. Specific verification data are provided to highlight the stepwise verification approach undertaken by a clinical diagnostic laboratory. Protocols for verification of in vitro diagnostic assays may vary between laboratories. However, all laboratories must verify several specific performance specifications prior to implementation of such assays for clinical use. We provide an example of an approach used for verifying performance of an assay for cystic fibrosis carrier screening.

Design and Analytic Validation of BCR-ABL1 Quantitative Reverse Transcription Polymerase Chain Reaction Assay for Monitoring Minimal Residual Disease

Monitoring minimal residual disease by quantitative reverse transcription polymerase chain reaction has proven clinically useful, but as yet there are no Food and Drug Administration-approved tests. Guidelines have been published that provide important information on validation of such tests; however, no practical examples have previously been published. To provide an example of the design and validation of a quantitative reverse transcription polymerase chain reaction test. To describe the approach used by an individual laboratory for development and validation of a laboratory-developed quantitative reverse transcription polymerase chain reaction test for BCR-ABL1 fusion transcripts. Elements of design and analytic validation of a laboratory-developed quantitative molecular test are discussed using quantitative detection of BCR-ABL1 fusion transcripts as an example. Validation of laboratory-developed quantitative molecular tests requires careful planning and execution to adequately address all required analytic performance parameters. How these are addressed depends on the potential for technical errors and confidence required for a given test result. We demonstrate how one laboratory validated and clinically implemented a quantitative BCR-ABL1 assay that can be used for the management of patients with chronic myelogenous leukemia.

UroVysion, Urine Cytology, and the College of American Pathologists: Where Should We Go From Here?

Abstract UroVysion (Abbott Molecular Inc, Des Plaines, Illinois) is a US Food and Drug Administration-approved test for the diagnosis of urothelial carcinoma. Although widely used, there are a variety of different ways to evaluate and interpret the test, and questions remain about the test's cost effectiveness and reproducibility in actual clinical practice. I suggest that the College of American Pathologists has a unique opportunity to create educational programs focusing on borderline or difficult urine cytology and UroVysion samples to provide data upon which to make evidence-based decisions concerning the best use of these tests.

Development of a Rapid Automated Influenza A, Influenza B, and Respiratory Syncytial Virus A/B Multiplex Real-Time RT-PCR Assay and Its Use during the 2009 H1N1 Swine-Origin Influenza Virus Epidemic in Milwaukee, Wisconsin

Rapid, semiautomated, and fully automated multiplex real-time RT-PCR assays were developed and validated for the detection of influenza (Flu) A, Flu B, and respiratory syncytial virus (RSV) from nasopharyngeal specimens. The assays can detect human H1N1, H3N2, and swine-origin (S-OIV) H1N1 Flu A viruses and were effectively used to distinguish Flu A infections (of all subtypes) from Flu B and RSV infections during the current S-OIV outbreak in Milwaukee, WI. The analytical limits of detection were 10−2 to 101 TCID50/ml depending on the platform and analyte and showed only one minor cross-reaction among 23 common respiratory pathogens (intermittent cross-reaction to adenovirus at >107 TCID50/ml). A total of 100 clinical samples were tested by tissue culture, both automated assays, and the US Food and Drug Administration-approved ProFlu+ assay. Both the semiautomated and fully automated assays exhibited greater overall (Flu A, Flu B, and RSV combined) clinical sensitivities (93 and 96%, respectively) and individual Flu A sensitivities (100%) than the Food and Drug Administration-approved test (89% overall sensitivity and 93% Flu A sensitivity). All assays were 99% specific. During the S-OIV outbreak in Milwaukee, WI, the fully automated assay was used to test 1232 samples in 2 weeks. Flu A was detected in 134 clinical samples (126 H1N1 S-OIV, 5 H1N1 [human], and 1 untyped) with 100% positive agreement compared with other “in-house” validated molecular assays, with only 2 false-positive results. Such accurate testing using automated high-throughput molecule systems should allow clinicians and public health officials to react quickly and effectively during viral outbreaks.

Re: Anti–Epithelial Cell Adhesion Molecule Antibodies and the Detection of Circulating Normal-Like Breast Tumor Cells

Identification of specific subtypes of circulating tumor cells in peripheral blood of cancer patients can provide information about the biology of metastasis and improve patient management. However, to be effective, the method used to identify circulating tumor cells must detect all tumor cell types. We investigated whether the five subtypes of human breast cancer cells that have been defined by global gene expression profiling-normal-like, basal, HER2-positive, and luminal A and B-were identified by CellSearch, a US Food and Drug Administration-approved test that uses antibodies against the cell surface-expressed epithelial cell adhesion molecule (EpCAM) to isolate circulating tumor cells. We used global gene expression profiling to determine the subtypes of a well-defined panel of 34 human breast cancer cell lines (15 luminal, nine normal-like, five basal-like, and five Her2-positive). We mixed 50-150 cells from 10 of these cell lines with 7.5 mL of blood from a single healthy human donor, and the mixtures were subjected to the CellSearch test to isolate the breast cancer cells. We found that the CellSearch isolation method, which uses EpCAM on the surface of circulating tumor cells for cell isolation, did not recognize, in particular, normal-like breast cancer cells, which in general have aggressive features. New tests that include antibodies that specifically recognize normal-like breast tumor cells but not cells of hematopoietic origin are needed.

Anti-Epithelial Cell Adhesion Molecule Antibodies and the Detection of Circulating Normal-Like Breast Tumor Cells

Identification of specific subtypes of circulating tumor cells in peripheral blood of cancer patients can provide information about the biology of metastasis and improve patient management. However, to be effective, the method used to identify circulating tumor cells must detect all tumor cell types. We investigated whether the five subtypes of human breast cancer cells that have been defined by global gene expression profiling—normal-like, basal, HER2-positive, and luminal A and B—were identified by CellSearch, a US Food and Drug Administration–approved test that uses antibodies against the cell surface–expressed epithelial cell adhesion molecule (EpCAM) to isolate circulating tumor cells. We used global gene expression profiling to determine the subtypes of a well-defined panel of 34 human breast cancer cell lines (15 luminal, nine normal-like, five basal-like, and five Her2-positive). We mixed 50-150 cells from 10 of these cell lines with 7.5 mL of blood from a single healthy human donor, and the mixtures were subjected to the CellSearch test to isolate the breast cancer cells. We found that the CellSearch isolation method, which uses EpCAM on the surface of circulating tumor cells for cell isolation, did not recognize, in particular, normal-like breast cancer cells, which in general have aggressive features. New tests that include antibodies that specifically recognize normal-like breast tumor cells but not cells of hematopoietic origin are needed.

Case Studies in Cost Effectiveness of Molecular Diagnostics for Infectious Diseases: Pulmonary Tuberculosis, Enteroviral Meningitis, and BK Virus Nephropathy

Pathogen genome amplification is used to detect and identify microorganisms, assess response to therapy, and detect mutations associated with drug resistance. Nucleic acid amplification tests have been shown to be superior to conventional culture-based testing methods in many circumstances. However, the enthusiasm for the technology in clinical laboratories may be decreased by the practical considerations of cost, complexity of the technology, and lack of US Food and Drug Administration-approved tests. The impact of nucleic acid amplification tests on the diagnosis and management of patients with tuberculosis, enteroviral meningitis, and BK virus transplant nephropathy will be examined, with an emphasis on the potential for health care cost savings.