Evaluation of Novel Polymerase Chain Reaction Method for the Diagnosis of Typhoid Fever in Suspected Cases

Ali  Amnan; Maria Mushtaq  Gill; Sakeenah Hussain  Naqvi; Adeel Hussain  Gardezi; Rabia  Sajjad; M Ali Khattak

doi:10.51253/pafmj.v76i4.12422

Authors

Ali Amnan Department of Microbiology, Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS), Rawalpindi Pakistan
Maria Mushtaq Gill Department of Microbiology, Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS), Rawalpindi Pakistan
Sakeenah Hussain Naqvi Department of Microbiology, Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS), Rawalpindi Pakistan
Adeel Gardezi Department of Microbiology, Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS), Rawalpindi Pakistan
Rabia Sajjad Department of Microbiology, Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS), Rawalpindi Pakistan
M Ali Khattak Department of Microbiology, Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS), Rawalpindi Pakistan

DOI:

https://doi.org/10.51253/pafmj.v76i4.12422

Keywords:

Blood culture, Diagnostic accuracy, Multiplex PCR, Enteric fever, Typhoid fever

Abstract

Objective: To evaluate viGen Q multiplex polymerase chain reaction (PCR) kit for typhoid fever using blood culture as the reference method in clinically suspected cases

Study Design: Diagnostic Accuracy Study

Place and Duration of Study: Department of Microbiology, Armed Forces Institute of Pathology Rawalpindi, Pakistan from Nov 2023 to Apr 2024.

Methodology: Venous samples of clinically suspected patients were collected in Ethylene Diamine Tetra Acetic Acid (EDTA) containing tube and blood culture bottle. On receipt in the Microbiology department of AFIP, a blood culture was performed and typhoidal salmonellae were identified to the species level by biochemical tests and serology. From the sample collected in the EDTA tube, deoxyribonucleic acid (DNA) was extracted using an automated DNA extractor (bioPerfectus), followed by amplification as per protocol of viGen Q multiplex PCR kit. Results were noted and data was analyzed using a statistical package for social sciences (SPSS) version 26.

Results: Blood samples of a total of 131 patients were tested by both blood culture and PCR methods for the detection of typhoidal salmonellae. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of viGen Q multiplex PCR kit were 92.7%, 94.4%, 88.4%, 96.6%, and 94% respectively. The Cohen’s kappa value (κ) was approximately 0.86, indicating strong agreement.

Conclusions: The viGen Q multiplex PCR kit has good sensitivity and specificity in comparison with blood culture and has the potential to expedite diagnosis and improve patient outcomes.

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