Can an Investigation of a Single Gene be Effective in Differentiating Certain Features of the Bipolar Disorder Profile?

Bipolar disorder (BD) is amongst the most common heritable mental disorders, but the clarification of its genetic roots has proven to be very challenging. Many single nucleotide polymorphisms (SNPs) have been identified to be associated with BD. SNPs in the CACNA1C gene have emerged as the most significantly associated with the disease. The aim of the present study is to provide a concise description of SNP 1006737 variants identified by Real Time PCR and confirm sequencing analysis with the Sanger method in order to estimate the association with BD. The molecular method was tested on 47 Sardinian subjects of whom 23 were found to not be mutated, 1 was found to be a carrier of the homozygous A allele and 23 were found to be carriers of the heterozygous G allele. Moreover, the positive results of the preliminary application suggest that the development of the screener could be extended to the other 5 genetic variables identified as associated with BD.

Reducing false-positive SARS-CoV-2 diagnoses using long-range RT-qPCR

Quantitative polymerase chain reaction (qPCR) is a sensitive molecular method for the detection of genetic material and regarded as the gold-standard for diagnostic testing. To detect respiratory RNA virus infections, a reverse transcription (RT) step is implemented to create cDNA molecules that can serve as template in the qPCR step. However, positive RT-qPCR results can be found long after patient recovery, in part because the RT-qPCR can detect residual viral RNA genome fragments. To minimize the detection of such fragments, we here modified the RT-qPCR assay by replacing the routinely used random hexamers with an oligonucleotide that binds to the 3' end of the viral genome. We demonstrate that this method allows us to distinguish between infectious and non-infectious samples. Moreover, in clinical samples obtained over 15 days after the onset of symptoms, we observe that the modified RT-qPCR protocol yields significantly fewer positive results compared to a commercial RT-qPCR test. No significantly different results were found compared to the commercial test when SARS-CoV-2 clinical samples were tested within 5 days of the onset of symptoms, suggesting that the modification has a similar sensitivity for detecting infectious viral RNA. Overall, these findings may help differentiate between incorrectly-positive, persistently positive, and reinfection cases in COVID-19 patients.

The Prevalence of Uropathogenic Escherichia coli Strains among Outpatients with Urinary Tract Infection in Zakho Hospitals-Zakho City, Duhok Province/ Iraq

This study involved the prevalence of uropathogenic Escherichia coli (UPEC) among outpatients of UTI attending three major hospitals in Zakho city. Four hundred urine samples were collected from patients of UTI of both sexes and different ages (≤ 1 year to over 50 years), during the period from July 2018 until January 2019. All urine samples were analyzed by conventional bacteriological method for the presence of Escherichia coli (E. coli), while molecular method was used for the presence of species-specific uidA gene in the isolated E. coli. Out of 400 samples, 141 (35.25%) were infected with UPEC from enrolled patients. The rate was higher in females than males (90.78% vs 9.22%), respectively. In both sexes, the age group 41-50 years in both sexes showed the highest rate (46.67%) of infection, and statistically this rate of infection was significant (p< 0.013) among both sexes and various age groups. Furthermore, in all ages, married patients showed slightly higher prevalence than un-married one (38% vs 32.5%), but this difference was statistically non-significant (p>0.05%). The rate of UTI was higher among urban inhabitants (40.56%) than others. During the months of the year, the peak (90.48%) in both sexes was during December while the lowest rates (13.64%) was during January.

Antimicrobial Spectrum, Growth/Killing Kinetics, Conventional/Molecular Assay of Characterizing Non-Leguminous Endophytic Bacteria and Fungi from Helianthus annuus, Carica papaya and Lycoperesicum solanum

The aim of this study is to comparative study between conventional and molecular assay of isolation, identification and characterization of non-leguminous endophytic bacteria and fungi in the leguminous root samples. The plant root samples, Helianthus annuus, Carica papaya and Lycoperesicum solanum (Sunflower root and stem, pawpaw root and stem, and tomato root and stem from Adekunle Ajasin University School farm, Akungba Akoko, Ondo state, Nigeria. The isolation of endophytic bacteria were performed using the conventional method of isolation (biochemical test) and characterization were done using both the conventional and molecular method of bacteria characterization. The antibiotic susceptibility test (Antibiogram) was observed using disc diffusion. The four bacteria identified were Bacillus cereus, Enterobacter sp. Actnomycoses sp. and Aeromonas sp. for conventional method and Fusarium solani, Fusarium vortecelium and Bacillus thuringiensis for molecular method as confirmatory point of view. In this study, all isolated organisms tends to be Gram positive using the gram staining technique. Antibiogram shows the zones of inhibition with diameter ranging from 0-20 mm, Enterobacter sp. were more sensitive to the various antibiotics used. Ultraviolet spectrophotometer was also used to determine the growth dynamic as well as the death rate of the isolates, the addition of antibiotics (ciprofloxacin) to the isolates at the 24th hour speed up the death rate of the isolates from non-leguminous endophytic bacteria. After the preliminary identification of the bacteria isolates and the confirmatory identification of both bacteria and fungi isolates of the non-leguminous endophytic microorganism, it was noted that the preliminary identification was only able to achieve the genus level of taxonomic characterization, While the molecular method confirm the molecular sub level identification of isolates depletes the absolute taxonomic identification and characterization to the sub-species level. The results of this study validates the use of molecular sequencing for the assay identification and characterization of non-leguminous endophytic bacteria and fungi as the easy and best mode of identification of both bacteria and fungi isolates as a veritable tools for research purposes.

Evaluation of the Different Methods to Detect Salmonella in Poultry Feces Samples

Salmonella is one of the most common causes of food-borne outbreaks and infection worldwide. The gold standard detection method of Salmonella is cultivation. With time-consuming cultivation, there is a need to investigate rapid and accurate processes. The study evaluated different approaches to detect Salmonella in poultry feces samples. Poultry farm feces samples from 21 cities in Iran were collected from January 2016 to December 2019. Microbiological cultures, serological assays, and multiplex PCR (m-PCR) were used to detect and characterize Salmonella spp. isolates. Serological assays and m-PCR were used to determine the serogroups A, B, C1, C2, D1, E, H, and FliC. The m-PCR was used for the detection of seven Salmonella serovars and a chi-square test was performed to compare the discriminatory power of the methods. Out of 2300 poultry feces samples, 173 (7.5%) and 166 (7.2%) samples were detected as Salmonella spp. by cultivation and m-PCR, respectively. The sensitivity of the molecular method was equal to cultivation at 0.96 (CI = 95%). Assessment of H antigenic subgroups showed the same for both m-PCR and serological tests. Therefore, the matching rate of the two methods for detection of all H antigenic subgroups was 100%. Thus, the relationship between the results obtained from both methods was significant in the contingency table test (P < 0.01). The PCR-based approach confirmed the detection of Salmonella in a shorter period (24–36 hours) compared to the conventional microbiological approach (3–8 days).