Jesús Tamariz1, Víctor Guevara2, Humberto Guerra3. 1. MSc, PhD, Antimicrobial resistance laboratory - Universidad Peruana Cayetano Heredia, Av. Honorio Delgado Nº 430, San Martín de Porras, Lima 31, Lima, Perú. 2. MT, Universidad Peruana Cayetano Heredia, Av. Honorio Delgado Nº 430, San Martín de Porras, Lima 31, Lima, Perú. 3. MD, PhD, Dr Med, Universidad Peruana Cayetano Heredia, Av. Honorio Delgado Nº 430, San Martín de Porras, Lima 31, Lima, Perú.
Abstract
INTRODUCTION: The salmonelloses are among the commonest, most widespread human zoonotic infections. They have generated international networks to attempt their control, since they cause a spectrum of ailments, ranging from inapparent carrier states to full-blown, severe, sometimes deadly diarrheal and systemic disease. Rapid diagnosis is needed for a number of reasons. The aim of this study was to standardize and validate a phage amplification test for the identification of salmonellosis to be applied to infections of Cavia porcellus. METHODS: Native bacteriophages were isolated from infected cavies and environmental residues from commercial cavy-breeding facilities. Salmonella enterica serovar Typhimurium ATCC 14028 was used to detect, isolate and propagate the bacteriophages, and to standardize a phage amplification assay to detect S. Typhimurium from rectal swabs of cavies. The phage amplification assay was tested using 2 antiviral agents, MgSO4·7H2O (MAS) and pomegranate rind extract (PRE) plus ferrous sulfate (PRE-FeSO4). RESULTS: The final assay format chosen used PRE-FeSO4 and allowed detection of S. Typhimurium in 90 min from culture, 5 h from clinical samples, with a limit of detection at 103 pfu; sensitivity was 98.2%, specificity 98%, negative predictive value (NPV) 96.1%, and positive predictive value (PPV) 99.1%. CONCLUSION: Bacteriophage amplification is therefore an appropriate, fast procedure for detection of this pathogen in clinical samples.
INTRODUCTION: The salmonelloses are among the commonest, most widespread human zoonotic infections. They have generated international networks to attempt their control, since they cause a spectrum of ailments, ranging from inapparent carrier states to full-blown, severe, sometimes deadly diarrheal and systemic disease. Rapid diagnosis is needed for a number of reasons. The aim of this study was to standardize and validate a phage amplification test for the identification of salmonellosis to be applied to infections of Cavia porcellus. METHODS: Native bacteriophages were isolated from infected cavies and environmental residues from commercial cavy-breeding facilities. Salmonella enterica serovar Typhimurium ATCC 14028 was used to detect, isolate and propagate the bacteriophages, and to standardize a phage amplification assay to detect S. Typhimurium from rectal swabs of cavies. The phage amplification assay was tested using 2 antiviral agents, MgSO4·7H2O (MAS) and pomegranate rind extract (PRE) plus ferrous sulfate (PRE-FeSO4). RESULTS: The final assay format chosen used PRE-FeSO4 and allowed detection of S. Typhimurium in 90 min from culture, 5 h from clinical samples, with a limit of detection at 103 pfu; sensitivity was 98.2%, specificity 98%, negative predictive value (NPV) 96.1%, and positive predictive value (PPV) 99.1%. CONCLUSION: Bacteriophage amplification is therefore an appropriate, fast procedure for detection of this pathogen in clinical samples.