Crizotinib efficacy in advanced non-squamous NSCLC patients with ALK or ROS1 rearrangement

AbstractIn patients with advanced non-small cell lung cancer (NSCLC), comprehensive genetic diagnostics is currently carried out in order to qualify for molecularly targeted therapies and immunotherapy. The aim of the study was to assess the usefulness of the reverse transcriptase (RT-PCR) method in the diagnosis of gene rearrangements, the effectiveness of EGFR, ALK, ROS1, and PD-L1 inhibitors in first-line treatment in NSCLC patients. We enrolled 95 non-squamous NSCLC patients with known status of EGFR, ALK, ROS1, MET and RET genes and PD-L1 protein expression. We used the real time PCR, fluorescence in situ hybridization (FISH), immunohistochemistry (IHC) and RT-PCR techniques for determination of predictive factors. In patients with ALK and ROS1 genes alteration, the median overall survival was 34 months in crizotinib treated patients and 6 months in patients who received chemotherapy (HR = 0.266, p = 0.0056). The risk of death was lower in patients treated with molecularly targeted therapies or immunotherapy compared to patients with predictive factors without personalized treatment (HR = 0.265, 95% CI 0.116–0.606) and to patient without predictive factors who received chemotherapy (HR = 0.42, 95% CI 0.162–1.09). Diagnosis of predictive factors and implementation of personalized treatment are key to prolonging the survival in advanced NSCLC patients.

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Tumor quantification of several fluoropyrimidines resistance gene expression with a unique quantitative RT-PCR method. Implications for pretherapeutic determination of tumor resistance phenotype

Cancer Letters ◽

10.1016/j.canlet.2005.11.008 ◽

2006 ◽

Vol 242 (2) ◽

pp. 168-179 ◽

Cited By ~ 4

Author(s):

Maud Barbado ◽

Laurence Preisser ◽

Michele Boisdron-Celle ◽

Veronique Verriele ◽

Gerard Lorimier ◽

...

Keyword(s):

Gene Expression ◽

Resistance Gene ◽

Tumor Resistance ◽

Rt Pcr ◽

Resistance Phenotype ◽

Pcr Method

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A highly specific real-time RT-PCR method for the quantitative determination of CK-19 mRNA positive cells in peripheral blood of patients with operable breast cancer

International Journal of Cancer ◽

10.1002/ijc.22017 ◽

2006 ◽

Vol 119 (7) ◽

pp. 1654-1659 ◽

Cited By ~ 59

Author(s):

Aliki Stathopoulou ◽

Maria Ntoulia ◽

Maria Perraki ◽

Stella Apostolaki ◽

Dimitris Mavroudis ◽

...

Keyword(s):

Breast Cancer ◽

Quantitative Determination ◽

Real Time ◽

Peripheral Blood ◽

Operable Breast Cancer ◽

Rt Pcr ◽

Pcr Method

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Comparison of a novel real-time PCR method (RTA) and Artus RG for the quantification of HBV DNA and HCV RNA

The Journal of Infection in Developing Countries ◽

10.3855/jidc.8363 ◽

2017 ◽

Vol 11 (07) ◽

pp. 543-548

Author(s):

Sibel Aydogan ◽

Ziya Cibal Acikgoz ◽

Aysegul Gozalan ◽

Fisun Kirca ◽

Tuba Muderris ◽

...

Keyword(s):

Quality Control ◽

Real Time ◽

Hbv Dna ◽

Clinical Samples ◽

Hcv Rna ◽

Rt Pcr ◽

B Virus ◽

Pcr Method ◽

Polymerase Chain

Introduction: The purpose of this study was to evaluate and compare two manual isolation and real-time (RT) polymerase chain reaction (PCR) kits (RTA RT-PCR with RTA isolation kit and Artus RG RT-PCR with QIAamp isolation kit) for molecular diagnosis of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections. Methodology: The study was conducted on 121 and 54 clinical samples for the detection of HBV DNA and HCV RNA, respectively, with an additional 8 HCV RNA external quality control samples. Results: Though a high correlation was observed between the two kits for the HBV DNA (r = 0.955, p = 0.001) and HCV RNA quantifications (r = 0.828, p = 0.001), discordant results were found in nine of the HBV DNA and in six of the HCV RNA samples. The mean difference between the two systems was found to be 0.4 log IU/mL in Quality Control for Molecular Diagnostics (QCMD) HCV RNA samples by Bland-Altman analysis. Conclusions: Although there was a high correlation between HBV DNA and HCV RNA tests according to the results of the study, the RTA system requires improvement for the determination of HCV RNA.

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METHOD FOR FMD VIRUS 146S COMPONENT CONCENTRATION DETERMINATION WITH REAL-TIME REVERSE TRANSCRIPTION – POLYMERASE CHAIN REACTION IN VACCINE RAW MATERIALS

Veterinary Science Today ◽

10.29326/2304-196x-2018-2-25-31-35 ◽

2018 ◽

pp. 31-35

Author(s):

M. I. Doronin ◽

A. M. Timina ◽

D. A. Lozovoy ◽

V. A. Starikov ◽

D. V. Mikhalishin ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Regression Model ◽

Real Time ◽

Reverse Transcription ◽

Raw Materials ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Method ◽

Polymerase Chain

Method for determination of foot-and-mouth disease (FMD) virus 146S component concentration with real-time reverse transcription – polymerase chain reaction (rtRT-PCR) in vaccine raw materials is developed. Negative regression model of С146S = (30.269 – Ct)/4.155 type allowing determination of FMDV 146S particle concentrations based on the amplification threshold cycle values (Ct) is proposed. It has been experimentally proven that the quantity of the 146S component determined by the real-time RT-PCR method using developed regression model and contained in the inoculation dose of FMD vaccine confers protection to the vaccinated animals against generalized FMD of A, O, Asia-1 types. rtRT-PCR method is highly sensitive and allows rapid and highly reliable estimation of the 146S antigen concentration in FMD vaccine. The method for 146S particle quantity determination by real-time RT-PCR using the regression model is reliable and demonstrates high correlation (95.5–99.0%) with the complement fixation test results.

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METHOD FOR FMD VIRUS 146S COMPONENT CONCENTRATION DETERMINATION WITH REAL-TIME REVERSE TRANSCRIPTION – POLYMERASE CHAIN REACTION IN VACCINE RAW MATERIALS

Veterinary Science Today ◽

10.29326/2304-196x-2018-2-25-26-30 ◽

2018 ◽

pp. 26-30 ◽

Cited By ~ 1

Author(s):

M. I. Doronin ◽

A. M. Timina ◽

D. A. Lozovoy ◽

V. A. Starikov ◽

D. V. Mikhalishin ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Regression Model ◽

Real Time ◽

Reverse Transcription ◽

Raw Materials ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Method ◽

Polymerase Chain

Method for determination of foot-and-mouth disease (FMD) virus 146S component concentration with real-time reverse transcription – polymerase chain reaction (rtRT-PCR) in vaccine raw materials is developed. Negative regression model of С146S = (30.269 – Ct )/4.155 type allowing determination of FMDV 146S particle concentrations based on the amplification threshold cycle values (Ct ) is proposed. It has been experimentally proven that the quantity of the 146S component determined by the real-time RT-PCR method using developed regression model and contained in the inoculation dose of FMD vaccine confers protection to the vaccinated animals against generalized FMD of A, O, Asia-1 types. rtRT-PCR method is highly sensitive and allows rapid and highly reliable estimation of the 146S antigen concentration in FMD vaccine. The method for 146S particle quantity determination by real-time RT-PCR using the regression model is reliable and demonstrates high correlation (95.5–99.0%) with the complement fixation test results.

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Development of a hemi-nested RT-PCR method for the specific determination of European Bat Lyssavirus 1

Vaccine ◽

10.1016/j.vaccine.2003.11.015 ◽

2004 ◽

Vol 22 (15-16) ◽

pp. 1921-1929 ◽

Cited By ~ 41

Author(s):

E Picard-Meyer ◽

V Bruyère ◽

J Barrat ◽

E Tissot ◽

M.J Barrat ◽

...

Keyword(s):

Rt Pcr ◽

Specific Determination ◽

Pcr Method

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Development of a method for the determination of the JAK2 gene mRNA in venous blood and assessment of its diagnostic value in oncohematology

Russian Clinical Laboratory Diagnostics ◽

10.51620/0869-2084-2021-66-6-379-384 ◽

2021 ◽

Vol 66 (6) ◽

pp. 379-384

Author(s):

M. A. Stolyar ◽

A. S. Gorbenko ◽

I. A. Olkhovskiy ◽

V. I. Bakhtina ◽

M. A. Mikhalev ◽

...

Keyword(s):

Multiple Myeloma ◽

Venous Blood ◽

Diagnostic Value ◽

Janus Kinase ◽

Rt Pcr ◽

Blood Samples ◽

Dnase Treatment ◽

Pcr Method ◽

Gene Mrna

Overactive JAK pathway signaling is a hallmark of immune diseases and critically affects on inflammation and coagulation. A number of mutations in the JAK2 gene act as driving forces of myeloproliferative neoplasms (MPN), the pathogenesis of certain variants of acute leukemia, a number of solid malignancies and cardiovascular diseases. Assays for quantifying JAK2 mRNA in circulating blood cells can be used as a marker associated with the activity of this enzyme. Development of an original method for detecting JAK2 mRNA in venous blood and assessment of the possible diagnostic value in chronic oncohematological diseases. The development of an RT-PCR method for determining the expression of the JAK2 gene mRNA in venous blood samples was carried out in accordance with the MIQE requirements. Primers and TaqMan probes were designed using the Primer3 program, taking into account the possibility of excluding subsequent DNase treatment. The stability of the investigated mRNA was assessed in vacutainers with different anticoagulants and depending on the storage time of the samples. The study of the expression of JAK2 mRNA in blood leukocytes of 41 patients with B-CLL, 16 patients with CML, 12 patients with multiple myeloma and 39 donors using the developed “real-time” PCR method. The study revealed a decrease in the level of JAK2 mRNA in venous blood samples in patients with primary CLL, but not with CML or with multiple myeloma. The level of the marker in the majority of patients with CLL after the start of therapy returned to the range typical for healthy people. It has been shown that the values of the relative expression of JAK2 mRNA are most stable in the range of 2 - 7 hours after taking blood in a vacutainer with EDTA. An original RT-PCR method was developed for the quantitative determination of JAK2 mRNA in venous blood samples, which meets the requirements of the MIQE system. Determination of JAK2 mRNA can be useful for clarifying the pathogenesis features of certain diseases involving impaired Janus kinase activity and can become a promising marker for prognosis and assessment of the effectiveness of therapy.

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