Use Of Molecular Tests Research Articles

An update on the Scianna blood group system

This update of the Scianna blood group system (Brunker PA, Flegel WA. Scianna: the lucky 13th blood group system. Immunohematology 2011;27:41-57) provides the recent work on the genetic variation of ERMAP across more world populations, the elucidation of the molecular basis of an historical serologic case, new cases of antibodies in the system, the development of new serologic reagents, and new discoveries in the biology of the erythroid membrane associated protein (ERMAP). Although genetic variation in ERMAP has been extensively cataloged, nonsynonymous variants associated with alloantigens have remained limited, and no new antigens have been identified. The first case of a severe hemolytic transfusion reaction to anti-Sc2 has recently been reported, highlighting the importance of pursuing the possibility of antibodies to low-prevalence antigens via indirect antiglobulin testing as a routine component of all transfusion reaction investigations. The expanding use of molecular testing in blood centers and transfusion services has uncovered a wider population distribution of Scianna antigens and heightened the awareness of this blood group system. The International Society of Blood Transfusion recognizes seven antigens in the Scianna blood group system 13.

Is Frozen-Section Analysis During Thyroid Operation Useful in the Era of Molecular Testing?

With the increased use of molecular testing of thyroid fine-needle biopsies, the frequency and extent of thyroid resection for thyroid nodules has changed. Although the role of frozen-section analysis of the thyroid has been reduced markedly in recent years, many surgeons still routinely use it intraoperatively. We sought to determine the utility of frozen section during thyroidectomy in the era of molecular testing. We reviewed 236 consecutive patients who had thyroidectomy with intraoperative frozen-section analysis at our institution between November 2015 and October 2017. We re-reviewed the preoperative diagnosis, frozen-section diagnosis, final pathology, and whether operative management changed from the initial plan based on frozen section. Mean age of the patients was 55.6 ± 14.1 years, and 83% were female. Of the 236 patients, frozen section did not change the intraoperative management in 225 (95%). Of the 11 patients whose thyroid operation was modified, the operation was either too much or not enough in 6 patients. In only 5 (2.1%) patients, frozen-section analysis correctly changed the extent of thyroidectomy. Thyroid frozen-section analysis adds cost and time to thyroid operations without notable benefit. In our cohort, only 2.1% of frozen sections accurately changed intraoperative management. We recommend against its routine use.

Porphyria cutanea tarda and hepatoerythropoietic porphyria: Identification of 19 novel uroporphyrinogen III decarboxylase mutations

Porphyria Cutanea Tarda (PCT) is a cutaneous porphyria that results from the hepatic inhibition of the heme biosynthetic enzyme uroporphyrinogen decarboxylase (UROD), and can occur either in the absence or presence of an inherited heterozygous UROD mutation (PCT subtypes 1 and 2, respectively). A heterozygous UROD mutation causes half-normal levels of UROD activity systemically, which is a susceptibility factor but is not sufficient alone to cause type 2 PCT. In both Types 1 and 2 PCT, the cutaneous manifestations are precipitated by additional factors that lead to generation of an inhibitor that more profoundly reduces hepatic UROD activity. PCT is an iron-related disorder, and many of its known susceptibility factors, which include infections (e.g. hepatitis C virus, HIV), high alcohol consumption, smoking, estrogens, and genetic traits (e.g. hemochromatosis mutations) can increase hepatic iron accumulation. Hepatoerythropoietic Porphyria (HEP) is a rare autosomal recessive disease that results from homozygosity or compound heterozygosity for UROD mutations and often causes infantile or childhood onset of both erythropoietic and cutaneous manifestations.During the 11-year period from 01/01/2007 through 12/31/2017, the Mount Sinai Porphyrias Diagnostic Laboratory provided molecular diagnostic testing for 387 unrelated patients with PCT and four unrelated patients with HEP. Of the 387 unrelated individuals tested for Type 2 PCT, 79 (20%) were heterozygous for UROD mutations. Among 26 family members of mutation-positive PCT patients, eight (31%) had the respective family mutation. Additionally, of the four unrelated HEP patients referred for UROD mutation analyses, all had homozygosity or compound heterozygosity for UROD mutations, and all eight asymptomatic family members were heterozygotes for UROD mutations. Of the UROD mutations identified, 19 were novel, including nine missense, two nonsense, one consensus splice-site, and seven insertions and deletions. These results expand the molecular heterogeneity of PCT and HEP by adding a total of 19 novel UROD mutations. Moreover, the results document the usefulness of molecular testing to confirm a genetic susceptibility trait in Type 2 PCT, confirm a diagnosis in HEP, and identify heterozygous family members.

Eye-Tracking Study to Enhance Usability of Molecular Diagnostics Reports in Cancer Precision Medicine.

We conducted usability studies on commercially available molecular diagnostic (MDX) test reports to identify strengths and weaknesses in content and form that drive clinical decision making. Given routine genomic testing in cancer medicine, oncologists must interpret MDX reports as well as evidence concerning clinical utility of biomarkers accurately for treatment or trial selection. This work aims to evaluate effectiveness of MDX reports in facilitating cancer treatment planning. Fourteen clinicians at an academic tertiary care medical facility, with a wide range of experience in oncology and in the use of molecular testing, participated in this study. Three commercially available, widely used, Clinical Laboratory Improvement Amendments (CLIA)-certified, College of American Pathologists (CAP)-accredited test reports (labeled Laboratories A, B, and C) were used. Eye tracking, surveys, and think-aloud protocols were used to collect usability data for these MDX reports focusing on ease of comprehension and actionability. Clinicians found two primary areas in molecular diagnostic reports most useful for patient care: therapy options with benefit or lack of benefit to patients, including enrolling clinical trials; and pathogenic tumor molecular anomalies detected. Therapeutic implications and therapy classes such as US Food and Drug Administration-approved off-label, on-label, clinical trials were critical for decision making. However, all reports had usability and comprehension issues in these areas and could be improved. Focused usability studies can help drive our understanding of the clinical workflow for use of molecular diagnostic tests in cancer care. This in turn can have major effects on quality of care, outcomes, costs, and patient satisfaction. This study demonstrates the use of specific usability techniques (eye tracking and think-aloud protocols) to help clinical laboratories improve MDX report design in a precision oncology treatment setting.

Acute hepatic porphyrias: Identification of 46 hydroxymethylbilane synthase, 11 coproporphyrinogen oxidase, and 20 protoporphyrinogen oxidase novel mutations

The acute hepatic porphyrias (AHPs) are inborn errors of heme biosynthesis, which include three autosomal dominant porphyrias, Acute Intermittent Porphyria (AIP), Hereditary Coproporphyria (HCP), and Variegate Porphyria (VP), and the ultra-rare autosomal recessive porphyria, δ-Aminolevulinic Acid Dehydratase Deficiency Porphyria (ADP). AIP, HCP, VP, and ADP each results from loss-of-function (LOF) mutations in their disease-causing genes: hydroxymethylbilane synthase (HMBS); coproporphyrinogen oxidase (CPOX); protoporphyrinogen oxidase (PPOX), and δ-aminolevulinic acid dehydratase (ALAD), respectively. During the 11-year period from January 1, 2007 through December 31, 2017, the Mount Sinai Porphyrias Diagnostic Laboratory diagnosed 315 unrelated AIP individuals with HMBS mutations, including 46 previously unreported mutations, 29 unrelated HCP individuals with CPOX mutations, including 11 previously unreported mutations, and 54 unrelated VP individuals with PPOX mutations, including 20 previously unreported mutations.Overall, of the 1692 unrelated individuals referred for AHP molecular diagnostic testing, 398 (23.5%) had an AHP mutation. Of the 650 family members of mutation-positive individuals tested for an autosomal dominant AHP, 304 (46.8%) had their respective family mutation. These data expand the molecular genetic heterogeneity of the AHPs and document the usefulness of molecular testing to confirm the positive biochemical findings in symptomatic patients and identify at-risk asymptomatic family members.

The burden of congenital Chagas disease and implementation of molecular diagnostic tools in Latin America

It is estimated that between 8000 and 15 000 Trypanosoma cruzi infected babies are born every year to infected mothers in Chagas disease endemic countries. Currently, poor access to and...

Descripción de pruebas moleculares en el diagnóstico del virus Zika en el periodo 2008-Febrero 2018. Revisión sistemática

Objetivo. Describir las pruebas moleculares usadas desde 2008 para la detección del virus Zika reportadas en artículos indexados en 3 bases de datos. Métodos. Se realizó la búsqueda de literatura científica utilizando el método PRISMA durante el periodo 2008 – febrero de 2018, en las bases de datos Pubmed, Science Direct y Embase, usando como criterios de inclusión artículos originales publicados entre los años 2008 a 2018 que realicen investigación en humanos con sospecha de infección por ZIKV y que describan la prueba molecular utilizada. Resultados. A partir de una búsqueda inicial de 2617 artículos recolectados con las palabras clave y aplicar los criterios de inclusión y exclusión, se seleccionan 18 artículos, de los cuales 8 son de Science Direct, 4 de Embase y 6 de Pubmed. Se describe la RT-PCR como la técnica más utilizada, se hallaron 9 blancos moleculares diferentes utilizados en las pruebas y 7 muestras biológicas de las cuales se pudo amplificar el genoma viral. Conclusiones. Hasta el momento la RT-PCR es la técnica más utilizada para el diagnóstico de la infección por el virus Zika al ser la más estandarizada y presentar una alta sensibilidad y especificidad, sin embargo se sigue trabajando sobre otras técnicas que garanticen la detección oportuna del virus, usando diferentes genes como el gen E, NS5, NS1, NS2B NS3, NS4B y CPREM principalmente en suero y orina.

Congenital erythropoietic porphyria and erythropoietic protoporphyria: Identification of 7 uroporphyrinogen III synthase and 20 ferrochelatase novel mutations

The erythropoietic porphyrias are inborn errors of heme biosynthesis with prominent cutaneous manifestations. They include autosomal recessive Congenital Erythropoietic Porphyria (CEP) due to loss-of-function (LOF) mutations in the Uroporphyrinogen III Synthase (UROS) gene, Erythropoietic Protoporphyria (EPP) due to LOF mutations in the ferrochelatase (FECH) gene, and X-Linked Protoporphyria (XLP) due to gain-of-function mutations in the terminal exon of the Aminolevulinic Acid Synthase 2 (ALAS2) gene. During the 11-year period from 01/01/2007 through 12/31/2017, the Mount Sinai Porphyrias Diagnostic Laboratory provided molecular diagnostic testing for one or more of these disorders in 628 individuals, including 413 unrelated individuals. Of these 628, 120 patients were tested for CEP, 483 for EPP, and 331 for XLP, for a total of 934 tests. For CEP, 24 of 78 (31%) unrelated individuals tested had UROS mutations, including seven novel mutations. For EPP, 239 of 362 (66%) unrelated individuals tested had pathogenic FECH mutations, including twenty novel mutations. The IVS3-48 T > C low-expression allele was present in 231 (97%) of 239 mutation-positive EPP probands with a pathogenic FECH mutation. In the remaining 3%, three patients with two different FECH mutations in trans were identified. For XLP, 24 of 250 (10%) unrelated individuals tested had ALAS2 exon 11 mutations. No novel ALAS2 mutations were identified. Among family members referred for testing, 33 of 42 (79%) CEP, 62 of 121 (51%) EPP, and 31 of 81 (38%) XLP family members had the respective family mutation. Mutation-positive CEP, EPP, and XLP patients who had been biochemically tested had marked elevations of the disease-appropriate porphyrin intermediates. These results expand the molecular heterogeneity of the erythropoietic porphyrias by adding a total of 27 novel mutations. The results document the usefulness of molecular testing to confirm the positive biochemical findings in these patients and to identify heterozygous family members.

Evidence behind the use of molecular tests in melanocytic lesions and practice patterns of these tests by dermatopathologists

The gold standard for the diagnosis of melanocytic lesions is histologic examination. However, as histologic examination can have its limitations, there are many clinical scenarios in which additional testing may be appropriate in an attempt to render a definitive diagnosis. A literature review for three ancillary tests-comparative genomic hybridization (CGH)/single-nucleotide polymorphism (SNP) array, fluorescence in situ hybridization (FISH), and gene expression profiling by quantitative reverse transcription polymerase chain reaction (qRT-PCR)-was compiled and current use patterns were tabulated. Survey of the practice patterns of these tests by dermatopathologists was also accessed in the attendees of the American Society of Dermatopathology Annual Meeting (Chicago, 2016). Here we summarize the use of these molecular tests in melanocytic lesions. We found that 54.4% of the respondents surveyed utilize (or expect consultants to utilize) molecular testing of melanocytic lesions in their practice when appropriate. CGH/SNP arrays, FISH testing, and qRT-PCR applied to melanocytic lesions have allowed for more accurate classification. Just over half of those surveyed use molecular testing for melanocytic lesion with the majority sending their cases out for completion of the molecular test.

Thyroid surgery for differentiated thyroid cancer - recent advances and future directions.

Population-based studies have demonstrated that an increasing number of incidental thyroid nodules are being identified. The corresponding increase in thyroid-based diagnostic procedures, such as fine-needle aspiration biopsy, has in part led to an increase in the diagnoses of thyroid cancers and to more thyroid surgeries being performed. Small papillary thyroid cancers account for most of this increase in diagnoses. These cancers are considered to be low risk because of the excellent patient outcomes, with a 5-year disease-specific survival of >98%. As a result, controversy remains regarding the optimal management of newly diagnosed differentiated thyroid cancer, as the complications related to thyroidectomy (primarily recurrent laryngeal nerve injury and hypoparathyroidism) have considerable effects on patient quality of life. This Review highlights current debates, including undertaking active surveillance versus thyroid surgery for papillary thyroid microcarcinoma, the extent of thyroid surgery and lymphadenectomy for low-risk differentiated thyroid cancer, and the use of molecular testing to guide decision-making about whether surgery is required and the extent of the initial operation. This Review includes a discussion of current consensus guideline recommendations regarding these topics in patients with differentiated thyroid cancer. Additionally, innovative thyroidectomy techniques (including robotic and transoral approaches) are discussed, with an emphasis on patient preferences around decision-making and outcomes following thyroidectomy.

Alternations in DNA gyrase genes in low-level fluoroquinolone-resistant Moraxella catarrhalis strains isolated in Poland.

PurposeThe purpose of this study was to investigate the molecular mechanisms of fluoroquinolone resistance in Moraxella catarrhalis clinical strains isolated in Lublin, Poland.Materials and methodsA total of 150 non-duplicate clinical strains of M. catarrhalis were obtained from individuals with signs of upper respiratory tract infection. Bacterial identification was corroborated on the basis of phenotypic and biochemical characteristics as well as with the use of molecular tests. The antimicrobial susceptibility of M. catarrhalis isolates was determined using the disk diffusion method and Etest. Mutations in the gyrase (gyrA and gyrB) and topoisomerase (parC and parE) genes were determined by polymerase chain reaction and sequencing.ResultsIt was observed that 16.7% of the studied isolates were drug resistant. Resistance to tetracycline was detected for 12% of the strains. Resistance to nalidixic acid, moxifloxacin, and levofloxacin was exhibited by 2.7% of the strains; 1.3% of the strains were resistant to trimethoprim/sulfamethoxazole and 0.7% to erythromycin. Minimum inhibitory concentration values of the four strains demonstrating fluoroquinolone resistance were: 6–12 mg/L for nalidixic acid, 1–1.5 mg/L for levofloxacin, 1 mg/L for moxifloxacin, and 0.25–0.5 mg/L for ciprofloxacin. The research resulted in the detection of mutations in 4 strains, in gyrase gyrA and gyrB genes. In gyrA gene, there occurred mutation G412C as well as four silent transition mutations. Within gyrB gene, there occurred mutation, substitution A1481G, as well as two identical silent mutations.ConclusionOur findings reveal that resistance to fluoroquinolones in M. catarrhalis is connected with amino acid substitutions in gyrA and gyrB genes. To our knowledge, this work is the first description of fluoroquinolone-resistant clinical strains of M. catarrhalis with described mutations in gyrA and gyrB genes isolated in Poland and in Europe.

Thyroid FNAC: Causes of false-positive results.

In this paper, we aim to focus on false positive results in the evaluation of thyroid aspirations, covering cystic, inflammatory, follicular and oncocytic lesions, papillary carcinoma, and medullary carcinoma of thyroid. The recently described entity noninvasive follicular thyroid neoplasm with papillary-like nuclear features is also discussed detailing the impact of its introduction on the sensitivity and specificity of thyroid FNA, as well as the use of molecular tests for diagnostics. Medicolegal issues in relation to current practice in English law are also described.

The 2016 WHO versus 2008 WHO Criteria for the Diagnosis of Chronic Myelomonocytic Leukemia.

The 2016 WHO diagnostic criteria for chronic myelomonocytic leukemia (CMML) require both absolute and relative monocytosis (≥1×109/L and ≥10% of white blood cell counts) in peripheral blood. Moreover, myeloproliferative neoplasm (MPN) features in bone marrow and/or MPN-associated mutations tend to support MPN with monocytosis rather than CMML. We assessed the impact of the 2016 WHO criteria on CMML diagnosis, compared with the 2008 WHO criteria, through a retrospective review of the medical records of 38 CMML patients diagnosed according to the 2008 WHO classification. Application of the 2016 WHO criteria resulted in the exclusion of three (8%) patients who did not fulfill the relative monocytosis criterion and eight (21%) patients with an MPN-associated mutation. These 11 patients formed the 2016 WHO others group; the remaining 27 formed the 2016 WHO CMML group. The significant difference in the platelet count and monocyte percentage between the two groups indicated that the 2016 WHO criteria lead to a more homogenous and improved definition of CMML compared with the 2008 WHO criteria, which may have led to over-diagnosis of CMML. More widespread use of molecular tests and more sophisticated clinical and morphological evaluations are necessary to diagnose CMML accurately.

Use of molecular testing and EGFR targeted medications for colorectal cancer at VA Hospitals.

e18814Background: Extended RAS testing in colorectal cancer predicts response to therapy with EGFR-directed therapies cetuximab and panitumumab (EGFR mab’s). In 2010 VA policy recommended testing o...

PMD6 - Real-World Data Analysis of Confirmatory Biopsy Utilization in Suspected Prostate Cancer Patients

As the US healthcare system transitions away from volume-based delivery to one that is focused on quality and value, there is a continued focus on understanding the drivers of cost, especially in oncology. Over one million prostate biopsies are performed annually, with many newly diagnosed cases being patients who may never die of the disease. Potential cost savings exist in identifying patients for which biopsy is unnecessary. In this study, we quantify the use of biopsy procedure in the diagnosis of prostate cancer. We utilized a chart pull survey methodology to identify 1,000 patients who had undergone traditional sextant prostate biopsy for suspected prostate cancer. Patient chart surveys were collected from 11 community urology sites varying in practice size and geographic representation from across the US. Patients were separated into research cohorts by whether they had received molecular testing prior to biopsy. Our research found that among the 1,000 patients who received a prostate biopsy, the most common reason for biopsy was elevated prostate-specific antigen (PSA), followed by abnormal digital rectal exam (DRE). Only 80 patients (8%) received a molecular test (4Kscore, PHI, PCA3, SelectMDx) prior to biopsy. Among the 1,000 patients who received a biopsy, 530 (53%) were confirmed to have prostate cancer. Among the 80 patients who had molecular testing prior to biopsy, 41 (51%) were confirmed to have prostate cancer. This result represents no significant difference in the rate of patients confirmed to have prostate cancer following biopsy (p=0.74), with or without prior molecular testing. Our results demonstrated that use of molecular testing did not improve the rate of prostate cancer confirmation through biopsy. They also highlighted the need for more accurate biomarkers for molecular testing to determine which patients should be biopsied among those suspected of having prostate cancer.

Outbreaks of diarrhoea ('winter scours') in weaned Merino sheep in south-eastern Australia.

Identify the cause of outbreaks of diarrhoea during winter that are not attributable to gastrointestinal nematodes in weaned Merino sheep in the high rainfall regions of south-eastern Australia and determine the efficacy of antimicrobials used to treat this syndrome. We investigated 45 outbreaks on 24 farms. Faecal samples from affected animals were cultured for Yersinia, Campylobacter and Salmonella spp. Risk factors, including rainfall, temperature and worm egg count (WEC), were assessed. Yersinia spp. were identified with molecular tests and susceptibility to four antimicrobials was determined. Yersinia pseudotuberculosis serotype III and virulent Y. enterocolitica were most frequently isolated. The frequency and severity of disease varied between region, farm and year. Yersinia pseudotuberculosis was detected only during winter, but Y. enterocolitica was present in all seasons. Pathogenic Yersinia species were more often isolated when WECs exceeded 500 eggs/g. A high proportion of Y. enterocolitica and Y. pseudotuberculosis were resistant to sulfafurazole (64% and 86.9%, respectively). A bacterial enteritis caused by pathogenic Yersinia was the cause of the winter scours syndrome in the 24 flocks investigated. The use of molecular testing increased the sensitivity of detection and identification of Yersinia spp. No clear association between weather, WEC and disease was established, suggesting complex interactions between risk factors are more important than any single factor. Sulfonamides should not be routinely used to treat this syndrome. Rather, during an outbreak the targeted use of an effective antimicrobial, such as oxytetracycline, should be integrated with grazing management strategies, including moving affected mobs onto lower risk pastures and decreasing the stocking rate.

Utilization of Molecular Testing and Survival Outcomes of Treatment with First- or Second-line Tyrosine Kinase Inhibitors in Advanced Non-small Cell Lung Cancer in a Dutch Population.

Epidermal growth factor receptor (EGFR) mutation testing is standard-of-care for advanced non-small cell lung cancer (NSCLC). Outcomes of second-/third-line compared to first-line tyrosine kinase inhibitors (TKIs) have shown conflicting results. We investigated utilization of molecular diagnostics and the outcomes of treatment with first-/second-line TKIs in patients with advanced NSCLC. Retrospective analysis was carried out of 2,206 patients with stage IIIb/IV NSCLC treated between 2008 and 2014 in four hospitals in the Netherlands. The rate of performing molecular diagnostics increased from 20.8% to 74.4% in the study period. The median overall survival of EGFR mutation-positive patients treated with TKIs was superior compared to EGFR mutation-negative patients treated with chemotherapy (720 vs. 274 days, p<0.0001). No difference in overall survival was found between EGFR mutation-positive patients treated only with TKIs compared to those treated with chemotherapy prior to TKIs, or upon progression under TKIs. The rate of EGFR testing has improved, increasing the number of patients eligible for targeted therapy. Chemotherapy, prior or subsequent to TKIs, for the treatment of EGFR mutation-positive patients, did not result in significantly better overall survival compared to that achieved with TKIs alone.

Genetic testing for indeterminate thyroid cytology: review and meta-analysis.

Thyroid cancer is the most frequent endocrine malignancy, and its incidence is increasing. A current limitation of cytological evaluation of thyroid nodules is that 20–25% are reported as indeterminate. Therefore, an important challenge for clinicians is to determine whether an indeterminate nodule is malignant, and should undergo surgery, or benign, and should be recommended to follow-up. The emergence of precision medicine has offered a valuable solution for this problem, with four tests currently available for the molecular diagnosis of indeterminate cytologies. However, efforts to critically analyze the quality of the accumulated evidence are scarce. This systematic review and meta-analysis is aimed to contribute to a better knowledge about the four available molecular tests, their technical characteristics, clinical performance, and ultimately to help clinicians to make better decisions to provide the best care options possible. For this purpose, we address three critical topics: (i) the proper theoretical accuracy, considering the intended clinical use of the test (rule-in vs rule-out) and the impact on clinical decisions; (ii) the quality of the evidence reported for each test (iii) and how accurate and effective have the tests proved to be after their clinical use. Together with the upcoming evidence, this work provides significant and useful information for healthcare system decision-makers to consider the use of molecular testing as a public health need, avoiding unnecessary surgical risks and costs.

WHO2016 Vs. WHO2008 Criteria for the Diagnosis of Chronic Myelomonocytic Leukemia

Abstract The new WHO2016 diagnostic criteria for chronic myelomonocytic leukemia (CMML) require both absolute and relative monocytosis (≥1×109/L and ≥10% of the WBC count) in peripheral blood (PB). In addition, they say that the presence of myeloproliferative neoplasms (MPN) features in the bone marrow (BM) and/or MPN-associated mutations (JAK2, CALR, or MPL) tend to support MPN with monocytosis rather than CMML. This study was performed to predict impact of the WHO2016 criteria in the diagnosis of CMML compared to the WHO2008 criteria. We retrospectively reviewed clinical and laboratory data of 38 patients diagnosed of CMML (32 CMML-1, 6 CMML-2) using the WHO2008 criteria in 7 university hospitals from Jan 2012 to Jul 2016. The laboratory data included complete blood cell count with WBC differentials, BM morphology, cytogenetic study, and molecular genetic study such as BCR-ABL1 and mutational status of JAK2 V617F, JAK2 exon12, CALR, and MPL . Male to female ratio was 2.8:1 and the mean age was 71±12 years. The medians of WBC count, hemoglobin, and platelet count were 22×109/L, 90 g/L and 154×109/L, respectively. The medians of monocyte, absolute monocyte count, blast, and nucleated red cell in PB were 20 (range 6~44)%, 4.1 (range 1.1~41.8)×109/L, 1 (range 0~10)%, and 0 (range 0~5)%, respectively. The medians of cellularity, M:E ratio, blast, and monocyte in BM were 90 (range 25~100)%, 5 (range 2~26), 3 (range 0~17)%, and 7 (range 1~26)%, respectively. MPN-associated mutations were present in 8 (7 JAK2 V617F, 1 MPL) out of 21 patients tested and BCR-ABL1 was negative in all 27 patients tested. Coefficient of utilization for MPN-associated mutations at diagnosis, was very low (3~11%) except for JAK2 V617F (50%). WHO2008 criteria classified the patients into 32 CMML-1 and 6 CMML-2. WHO2016 criteria excluded 3 patients for not fulfilling relative monocytosis and 8 patients for having a MPN-associated mutation, and classified the 27 CMML patients into 17 CMML-0, 5 CMML-1, and 5 CMML-2. Sixteen was proliferative type (WBC count≥13×109/L) of CMML and 11 dysplastic type (WBC count The relative monocytosis (≥10% of the WBC count) was not met in 3 (8%) patients and MPN-associated mutations were found in 8 (21%) patients. The WHO2008 criteria might over diagnose CMML. More aggressive use of molecular tests including MPN-associated mutation analyses and more sophisticated clinical or morphologic evaluation are necessary in order to make an accurate diagnosis of CMML. Disclosures No relevant conflicts of interest to declare.

P2.03-032 Efficacy and Safety of Osimertinib as Third-Line or Later Therapy for T790M-Positive Advanced Non-Small Cell Lung Cancer

Advanced lung adenocarcinoma with epidermal growth factor receptor (EGFR) gene mutation in exon 18 to 21 is sensitive to EGFR tyrosine kinase inhibitors (TKI) such as gefitinib, erlotinib, and afatinib. However acquired resistance to EGFR-TKI develops after initial treatment, primarily due to T790M mutation. In March 2016, the Japanese Ministry of Health, Labor and Welfare approved osimertinib for the patients with advanced non-small cell lung cancer harboring a T790M mutation. In the same year, patients with a T790M secondary mutation were reviewed at Hiratsuka Kyosai Hospital for a period of 9 months. All patients were previously treated with an EGFR-TKI (with or without or prior/subsequent chemotherapy). Data were obtained retrospectively by analysis of the medical records of patients who underwent molecular diagnostic testing for T790M mutation. All patients had stage IV adenocarcinoma. A total of 8 patients with T790M mutation were identified. Molecular testing was performed using re-biopsy specimens of the primary tumor, metastatic lymph node, or circulating DNA in the plasma of patients. Seven out of 8 patients received osimertinib 80mg once daily. All patients treated with osimertinib received other treatments previously, including EGFR-TKI and standard chemotherapy. The overall response rate (ORR) was 87%. Most common adverse events included diarrhea (28.6%), rash (14.2%), nausea (14.2%). Adverse events of Grade 3 to 4 severity were not reported. These findings suggest that third-line or later osimertinib for advanced lung adenocarcinoma with T790M results in high ORR and managed tolerability. The use of molecular testing may improve treatment outcome in these patients.