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A Rapid and Sensitive One Step-SYBR Green Based Semi Quantitative Real Time RT-PCR for the Detection of peste des petits ruminants Virus in the clinical samples

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Virologica Sinica 2012年第1期27卷 1-9页

作者： Vinayagamurthy Balamurugan Arnab Sen Gnanavel Venkatesan Vinita Yadav Vandna Bhanot Veerakyathappa Bhanuprakash Raj Kumar Singh Division of Virology Indian Veterinary Research InstituteMukteswar 263138UttarakhandIndia Project Directorate on Animal Disease Monitoring And Surveillance(PD-ADMAS) Bangalore 560024KarnatakaIndia Animal Health Division ICAR-NEH RegionMeghalaya 793103India Indian Veterinary Research Institute Bangalore 560024KarnatakaIndia National Research Centre on Equines Hisar 125001HaryanaIndia

A sensitive and rapid single step real time （rt） RT-PCR was standardized using one-step Brilliant SYBR Green kit for detection and semi-quantitation ofpeste des petitis ruminants virus （PPRV） using the virus RNA and matrix （M） protein gene-specific primers and compared with established conventional RT-PCR and TaqMan RT-PCR. The assay amplifies a 124 bp fragment of the PPRV M gene with Tm of 78.28 to 78.50. The assay was linear within a range of 50 ng to 0.5 fg total virus RNA with a detection limit （sensitivity） of 0.5 fg. Based on the serial dilution of the live-attenuated PPR vaccine virus, the detection limit was -0.0001 cell culture infectious dose 50% units （TCID50）. Additionally, swab materials spiked with known titre of vaccine virus were equally well detected in the assay. The standardized rt RT-PCR was easily employed for the detection of PPRV nucleic acid directly in the field and experimental clinical samples. The assay detected the PPRV nucleic acid as early as 3 day post infection （dpi） and up to 20 dpi in swab materials from the experimental samples. The assay was rapid and more sensitive than TaqMan and conventional RT-PCR in the detection of PPRV nucleic acid from the PPR suspected clinical samples of sheep and goats. Therefore, the established, simplified SYBR green rt RT-PCR is an alternative test to the already existing various diagnostic assays and could be useful for rapid clinical diagnosis with advantage in reducing risk of contamination.

关键词： PPR M gene SYBR green RT-PCR Early diagnosis clinical samples

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Assessment of Appropriateness of Diagnostic Lymph Node Tissue Collection from the Operating Theater and by Interventional Radiologist. A clinical Audit

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Advances in Infectious Diseases 2024年第3期14卷 541-548页

作者： Kowthar Salman Hassan Divya Deodhar Abdullah T. Al-Rawahi Mahmood H. Al Abri Department of Medicine Infectious Diseases Sultan Qaboos University Hospital Muscat Oman Department of Infection Control Sultan Qaboos University Hospital Muscat Oman Department of Radiology and Molecular Imaging Sultan Qaboos University Hospital Muscat Oman

Background: Appropriate sample requesting, collecting and timely dispatch to the appropriate laboratory is essential in establishing diagnosis of pathologies with lesions. Much time and effort may be wasted if this is not done according to certain standards. We conducted this study to assess the route of lymph node samples from requests to reaching the laboratories. Methods: We conducted an audit over a period from 4th June until 10th Aug 2023. Data for all the procedures performed over this period on lymph node samples (was entered into and analysed using Excel. Results: A total of eighteen samples for sixteen patients were obtained during this period. Median age of the patients was 34 years (19 - 73) with a M:F ratio of 5:11. Among the IR samples, nine samples were from the neck, three from inguinal area and one from axilla. Seven samples (53.8%) were tru-cut biopsies, six samples (46.15%) were FNA. All samples were sent to the pathology laboratory fixed in formalin. samples for TB were sent only for five cases (31.25%) and for only two cases (12.5%) were samples sent for bacterial culture. For the OR samples, none were sent for either bacterial culture or TB. Overall, eight patients (50%) were not investigated for any infectious etiologies like brucella, toxoplasmosis, CMV, EBV plus other possible causes. Repeat sampling was required for 25% of patients (within and out of the audit period). Conclusions: to avoid delays in making diagnoses, it is paramount to consider infectious etiologies as possible diagnosis for lymphadenopathy and request appropriate investigations. This requires liaising with infectious diseases/clinical microbiology experts to guide regarding types of samples, types of media and timely dispatch to the correct laboratory.

关键词： clinical Audit clinical samples Biopsies Lymph Nodes Saline Formalin Quality Improvement

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