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September 2021 Volume 7 Issue 3

“Evaluation of Different Commercially Available Real Time-Polymerase Chain Reaction Kits for SARS-CoV-2 Diagnosis”

Suneet kr. Yadav, Dr. R.Sujatha, Deepak Sameer bind

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Introduction: - At present the whole world is facing pandemic of the Corona virus disease (COVID-19) caused by severe acute respiratory syndrome corona virus 2 (SARS-CoV-2). The knowledge of available laboratory methods is essential for early and correct diagnosis of COVID-19 to identify new cases as well as monitoring treatment of confirmed cases.

Aim: - To compare basic analytical and clinical performance of selected RT-PCR kits from Three different manufacturers ( LaboGun, Gene Store, TruPCR).

Materials and Methods: Three commercially available kits have been evaluated on the basis of: (i) number of SARS-CoV-2 specific gene target; (ii) human housekeeping genes as internal control; (iii) RT-PCR run time; and (iv) kit performances to correctly detect SARS-CoV- 2 positive and negative RNA samples. A total of 50 RNA samples (left over RNA) were included, master mix preparation, template addition and RT-PCR test has been performed according to kits literature. At the end of PCR run, mean and standard deviation of obtained cut- off of all kits were calculated using Microsoft Excel.

Result: In this study all three RT-PCR kits performed satisfactory regarding the reproducibility and they could correctly identify RNA samples. RNA samples having low viral loads with a high Cycle threshold (Ct) value (>30) were also detected by all these three kits. Obtained Ct values of each group was in parallel range in comparison with the initial testing Ct values. Kits were found to be superior who contains primers and probes for three SARS-CoV-2 specific gene targets, have human housekeeping gene as internal control and taking less time to complete RT-PCR.

Conclusion: - All three COVID-19 RT-PCR kits included in this study demonstrated satisfactory performance and can be used for the routine molecular diagnosis of COVID-19 disease.


“To Study the Microbiological Profile and MDR Strains of Chronic Non Healing Diabetic Ulcers with Special Reference to Bio film Formation”

Nashra Afaq, R. Sujatha

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Background: Diabetes mellitus is a major health problem that affects approximately 171 million people globally. One of its most severe complications is the development of diabetic foot ulcers (DFU). Multidrug Resistance (MDR) is basically caused by the Bio film which developers a barrier for the immune system and the antimicrobial agents.

Aim and Objective: To Study the Microbiological Profile and MDR Strains of Chronic Non Healing Diabetic Ulcers with Special Reference to Bio film Formation.

Material and Methods: This is a cross-sectional study which was carried out in the Department of Microbiology, RMCH &RC, Mandhana, and Kanpur for a period of 1 year January 2020 to December 2020. Clinical samples from Pus and tissue bit samples were taken from 200 diabetic patients with non healing ulcers as per standard microbiological procedures. Antimicrobial susceptibility test was performed as per Clinical and Laboratory Standards Institute (CLSI) guidelines. Detection of Bio film formation by the tissue culture plate method was done.

Results: Out of 200 diabetic patients, the ratio of Males were more than the Females with 122 (61%) and 78 (39%). Patients with type 2 diabetes related ulcer were more than Type 1.  The most common sites of ulcers were the plantar surface of foot 101 (50.5%), toes 49 (24.5%), and dorsal surface of foot 50 (25%). Among the gram negative aerobes 120 (60%) are predominant than gram positive aerobes 47 (23.5%). The rest of the growth showed 22 (11%) Fungal and 11 (5.5%) anaerobic growth.

 The predominantly isolated pathogens were Pseudomonas aeruginosa and Staphylococcus aureus among aerobic bacteria, Peptostreptococcus among the anaerobes and Candida alb cans was most predominantly isolated among fungus.

Staphylococcus aureus was the strong biofilm producer, followed by Pseudomonas aeruginosa, baumannii. Gram-negative bacteria showed high sensitivity to piperacillin-tazobactam, meropenem, Coniston, gram-positive cocci to vancomycin and linezolid .

Conclusion: Bio films and polymicrobial infection have a crucial role in DFIs and contribute to delay healing. These wounds are characterized by a complex micro biome and a polymicrobial organization, especially within the bio film. The development of processes and methodologies to study bio films is needed. This represents the next step to validating new anti bio film molecules with a promising therapeutic potential.

“Evaluation of Rapid Antigen Test (RAT) and Real‑Time Polymerase Chain Reaction (RT-PCR) for detection of COVID‑19 in Kanpur UP”

Suneet kr. Yadav, R. Sujatha, Deepak Sameer

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Introduction: Severe acute respiratory syndrome corona virus 2 (SARS-CoV-2), which emerged as a novel human pathogen in China at the end of 2019, is responsible for corona virus disease 2019 (COVID-19), which causes symptoms such as cough and fever, severe pneumonia, and death. The WHO reported that more than 29 million cases of COVID-19, including approximately 55, 40,000 deaths, have occurred as of 17 January 2022

Aim: To Evaluation of Rapid Antigen Test (RAT) and Real‑Times Polymerase Chain Reaction (RT-PCR) for detection of COVID‑19 in kanpur UP.

Materials and Methods: This study was be conducted in the Department of Microbiology Rama Medical College Hospital and Research Centre Kanpur. Total 100 known samples, out from 50 positive by RAT patient and 50 negative patients already tested by RAT kit for Covid -19 test. For the present study, Tru PCR SARS-CoV-2 RT-q PCR Kit (Kilpest India Ltd., India) were selected on the basis of multiple SARS-CoV-2 specific gene and Standard Q COVID-19 Ag kit (SD Biosensor, Healthcare Pvt. Ltd Guru gram Haryana India) is a rapid chromatographic immunoassay for the detection of SARS-CoV-2 nucleocapsid (N) antigen in respiratory specimen.

Result: In this study100 respiratory samples collected from individuals living in a shared housing were analyzed head to head by Rapid antigen test and RT-PCR using CFX-96 real-time thermal cycler. Out of 100, 50 negative samples by RAT, 21 (10.5%) of the  samples were found positive by SARS-CoV-2 by rRT-PCR with cT values ranging between 17.32–32.91 and 50 positive samples by RAT, they were all 50 (100%) samples found positive by SARS-CoV-2 by rRT-PCR with cT values ranging between 16.62–33.91 The antigen test diagnosed the infection status with a sensitivity of 79.0 % (79/100) and a specificity of 100 %.

Conclusion: All six COVID-19 RT-PCR kits included in this study demonstrated satisfactory performance and can be used for the routine molecular diagnosis of COVID-19 disease

Prevalence of Co-Infection with Salmonella Typhi and Malaria Parasites in Tertiary Care Centre Kanpur

Deepak Sameer, R. Sujatha, Suneet Kr Yadav, Sarah K

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Background: Malaria and typhoid fever often present with mimicking symptoms especially in the early stages of typhoid On the other hand, typhoid fever is also a major public health problem in India. It is an acute systemic infection caused by the bacterium Salmonella Typhi exaggerates the situation. Thus the aim of this work was to investigate the rate of co-infection with respect to the use of widal test and blood culture methods for diagnosing typhoid fever in Kanpur.

Methods: This study was conducted in Rama Medical College, Hospital & Research Centre, and Kanpur. It is a retrospective study conducted from July 2021 to December 2021 A total of 654 blood samples were collected (5ml of blood drawn by vene puncture) from each febrile patients, both OPD and IPD, who were tested for widal test and Malaria card test for malaria parasite detection. Patients were explained about the study and their consent was taken.

Results: The results of this study are based on bacteriological and serological tests for the diagnosis of typhoid fever parasitological examination for malaria parasites and in 126 patients.

The total 654 sample where 126 samples were positive for typhoid by the widal test and 27 (21.4%) (Fig:-2) samples were positive for malaria parasites by card, only 6 by the culture method. The rate of co-infection was significantly high when typhoid was diagnosed by widal (19.26%) than by blood culture method (4.7%). The patients comprised 78 (61.9%) females and 48 (38%) males (fig:-1) aged between 14 to 65 years (mean= 41 years) Malaria parasites were found in 27 (21.4%) samples henceforth known as malaria patients.

Conclusion: The incidence of typhoid and malaria co-infection will greatly reduce if the diagnosis of typhoid fevers in malaria endemic area.


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