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The current study attempts the phenotypic and molecular detection of the virulence factors of Proteus mirabilis clinical isolates. A total of 600 urine samples were collected from urinary tract-infected patients at Khyber Teaching Hospital, Peshawar, Pakistan. P. mirabilis isolates were identified through different biochemical tests. Virulence factors including urease production, biofilm formation, and swarming phenomenon were determined by using different markers (ureC1, rsbA, and luxS genes), identified via the polymerase chain reaction (PCR) technique. Out of the selected samples, 95 samples (15.83%) were identified as P. mirabilis. The molecular study showed that all isolates (100%) possessed the ureC gene. Whereas, 90.52% and 92.63% of the isolates gave positive results for biofilm formation (rsbA gene) and swarming phenomenon (luxS gene), respectively. The phenotypic and molecular study of P. mirabilis virulence factors provides a better understanding of how P. mirabilis infection spreads. The results could be used for prevention and improvements in its clinical treatment.

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Myeloproliferative Neoplasms (MPNs) are rare heterogeneous hematological disorders usually characterized by one or more lineages of myeloid cells in bone marrow and increase number of normal and abnormal cells. Janus kinase 2 valine to phenylalanine (JAK2-V617F) is usually present in Philadelphia-negative MPNs. Pathogenic mutation in JAK2-V617F cause’s valine to phenylalanine substitution in JAK2 gene on exon-14. Different methods such as Allele-specific PCR (AS-PCR), Amplification refractory mutation system (ARMS-PCR), High resolution melting (HRM) analysis and Molecular beacon probe-based RT-PCR are already available to diagnose JAK2-V617F mutation. In current study, we aimed to develop and optimize real-time PCR assay which will be available locally and be feasible, less expensive and less labor extensive. The DNA was extracted from 128 patients and analyzed on our optimized method using newly designed primers and probe. Standards were generated using in-vitro synthesized sequence (Kinco Biological) and Standard curve was obtained. Predicted sensitivity of the method is at least5% for allele burden of the mutation. The total of 128 MPN patients were included in the present study and 54 (42.1%) were JAK2-V617Fpositive according to the optimized protocols. The study concluded that TaqMan Real time PCR is sensitive, efficient and less expensive for the detection of JAK2-V617F mutation.