Molecular screening of Bartonella in free-ranging capybaras (Hydrochoerus hydrochaeris) from Paraná State, Southern Brazil

Bartonella is an emerging group of facultative intracellular bacteria causing circulatory and systemic disorders. Hosts for Bartonella are mostly mammals, specifically rodents, having a growing number of Bartonella species related to their infection. Capybaras (Hydrochoerus hydrochaeris) are abundant native rodents of Brazil, commonly found in urban parks. In the present study, we aimed to perform molecular screening of capybaras for Bartonella spp. Blood samples were collected from 17 free-ranging animals captured in Paraná State, Southern Brazil. None of the collected samples tested positive for the Bartonella-nuoG gene by quantitative polymerase chain reaction (qPCR), although all of them successfully amplified the mammal endogenous glyceraldehyde-3-phosphate dehydrogenase (gapdh) gene. Additionally, all animals were infested exclusively by Amblyomma dubitatum ticks at the time of sampling. This study was part of an active surveillance program, which is critical for monitoring animal health status, particularly in capybaras.

LUMOS - Low and Intermediate Grade Glioma Umbrella Study of Molecular Guided TherapieS at relapse: Protocol for a pilot study

IntroductionGrades 2 and 3 gliomas (G2/3 gliomas), when combined, are the second largest group of malignant brain tumours in adults. The outcomes for G2/3 gliomas at progression approach the dismal outcomes for glioblastoma (GBM), yet there is a paucity of trials for Australian patients with relapsed G2/3 gliomas compared with patients with GBM. LUMOS will be a pilot umbrella study for patients with relapsed G2/3 gliomas that aims to match patients to targeted therapies based on molecular screening with contemporaneous tumour tissue. Participants in whom no actionable or no druggable mutation is found, or in whom the matching drug is not available, will form a comparator arm and receive standard of care chemotherapy. The objective of the LUMOS trial is to assess the feasibility of this approach in a multicentre study across five sites in Australia, with a view to establishing a national molecular screening platform for patient treatment guided by the mutational analysis of contemporaneous tissue biopsiesMethods and analysisThis study will be a multicentre pilot study enrolling patients with recurrent grade 2/3 gliomas that have previously been treated with radiotherapy and chemotherapy at diagnosis or at first relapse. Contemporaneous tumour tissue at the time of first relapse, defined as tissue obtained within 6 months of relapse and without subsequent intervening therapy, will be obtained from patients. Molecular screening will be performed by targeted next-generation sequencing at the reference laboratory (PathWest, Perth, Australia). RNA and DNA will be extracted from representative formalin-fixed paraffin embedded tissue scrolls or microdissected from sections on glass slides tissue sections following a review of the histology by pathologists. Extracted nucleic acid will be quantified by Qubit Fluorometric Quantitation (Thermo Fisher Scientific). Library preparation and targeted capture will be performed using the TruSight Tumor 170 (TST170) kit and samples sequenced on NextSeq 550 (Illumina) using NextSeq V.2.5 hi output reagents, according to the manufacturer’s instructions. Data analysis will be performed using the Illumina BaseSpace TST170 app v1.02 and a custom tertiary pipeline, implemented within the Clinical Genomics Workspace software platform from PierianDx (also refer to section 3.2). Primary outcomes for the study will be the number of patients enrolled and the number of patients who complete molecular screening. Secondary outcomes will include the proportion of screened patients enrolled; proportion of patients who complete molecular screening; the turn-around time of molecular screening; and the value of a brain tumour specific multi-disciplinary tumour board, called the molecular tumour advisory panel as measured by the proportion of patients in whom the treatment recommendation was refined compared with the recommendations from the automated bioinformatics platform of the reference laboratory testing.Ethics and disseminationThe study was approved by the lead Human Research Ethics Committee of the Sydney Local Health District: Protocol No. X19-0383. The study will be conducted in accordance with the principles of the Declaration of Helsinki 2013, guidelines for Good Clinical Practice and the National Health and Medical Research Council National Statement on Ethical Conduct in Human Research (2007, updated 2018 and as amended periodically). Results will be disseminated using a range of media channels including newsletters, social media, scientific conferences and peer-reviewed publications.Trial registration numberACTRN12620000087954; Pre-results.

Molecular Screening of Some Apple Progenies for Detection Vf Gene Using Marker Assisted Selection

Apple scab, caused by Venturia inaequalis is one of the most damaging pathogens that affects apple species. Cross combinations were made between Auriu de Bistrița cv. (female genitor) - a valuable local variety in terms of fruit quality but only tolerant to scab, and Florina cv. (male genitor) used as a donor of Vf resistance gene. It was first detected in Malus floribunda Clone 821, which was later transferred to commercial varieties by different breeding programs. To confirm the presence of Vf gene, progenies resulting from the mentioned combination were tested with MAS (Marker Assited Selection), using two dominant primers pairs (AM19, U1400), and another one codominant (AL07) used to distinguish homozygous and heterozygous genotypes. After the crossing combination, a number of twenty-six hybrids were obtained, of which 50% (13 hybrids) were clasified as resistant (heterozygous), respectively 50% (13 hybrids) as susceptible (recessive homozygotes), so the Mendelian ratio was confirmed.

First Nationwide Molecular Screening Program in Spain for Patients With Advanced Breast Cancer: Results From the AGATA SOLTI-1301 Study

BackgroundThe SOLTI-1301 AGATA study aimed to assess the feasibility of a multi-institutional molecular screening program to better characterize the genomic landscape of advanced breast cancer (ABC) and to facilitate patient access to matched-targeted therapies in Spain.MethodsDNA sequencing of 74 cancer-related genes was performed using FFPE tumor samples in three different laboratories with three different gene panels. A multidisciplinary advisory board prospectively recommended potential targeted treatments. The primary objective was to determine the success of matching somatic DNA alteration to an experimental drug/drug class.ResultsBetween September 2014 and July 2017, 305 patients with ABC from 10 institutions were enrolled. Tumor sequencing was successful in 260 (85.3%) patients. Median age was 54 (29-80); most tumors were hormone receptor-positive/HER2-negative (74%), followed by triple-negative (14.5%) and HER2-positive (11.5%). Ninety-seven (37%) tumor samples analyzed proceeded from metastatic sites. Somatic mutations were identified in 163 (62.7%) patients, mostly in PIK3CA (34%), TP53 (22%), AKT1 (5%), ESR1 (3%), and ERBB2 (3%) genes. Significant enrichment of AKT1 mutation was observed in metastatic versus primary samples (9% vs. 2%; p=0.01). Genome-driven cancer therapy was recommended in 45% (n=116) of successfully screened patients, 11% (n=13) of whom finally received it. Among these patients, 46.2% had a PFS of ≥6 months on matched therapy.ConclusionsAGATA is the first nationwide molecular screening program carried out in Spain and we proved that implementing molecular data in the management of ABC is feasible. Although these results are promising, only 11% of the patients with genome-driven cancer therapy received it.

Serological and Molecular Screening of Camels for Brucellosis in Bikaner, Rajasthan, India

Brucellosis is an important zoonotic disease affecting domestic animals and humans worldwide. The present study was undertaken on camels in and around Bikaner city of Rajasthan state of India to assess the extent of prevalence of Brucellosis in camels in this region. Since Rose Bengal Plate test (RBPT) is a serological screening method for diagnosis of Brucellosis approved by the Office International des Epizooties (OIE), RBPT was employed for detecting antibodies against Brucella organisms in camels. Polymerase Chain Reaction is widely followed for molecular diagnosis of several infectious diseases. DNA from whole blood of camels was analyzed by PCR for detection of Brucella organisms in the blood of camels. Blood samples from 177 camels (108 males and 69 females) from Bikaner and nearby villages were analyzed for Brucellosis by RBPT. Fifteen camels [7 (46.66%) males and 8 (53.33%) females] were found positive. However, none of the DNA samples from whole blood (RBPT positive or negative) from 25 camels tested was Brucella positive by PCR. The serological results indicate that Brucellosis is prevalent in camels and is of public health significance in Bikaner and nearby villages in Rajasthan state of India. However, detection of DNA of Brucella organisms in blood by PCR may not be advised for regular screening for Brucellosis since there is intermittent bacteremia in Brucellosis and Brucella DNA may not be detectable in blood continuously throughout the course of the disease. This reminds us that the OIE has approved RBPT, but not PCR for screening of Brucellosis.