New Flagellin Gene for Salmonella enterica serovar Typhi from the East Indonesian Archipelago

Abstract

Phase variation is a property unique of some Salmonella enterica serovar Typhi strains from Indonesia. Salmonella Typhi isolates from Indonesia have been described that in addition to the phase 1 Hd flagellin gene contain a second flagellin gene named z66. S. Typhi isolates from Indonesia with a mutant Hd gene named Hj have also been described. Here, we have identified another flagellin gene of S. Typhi, named Ind, showing a closest homology with the flagellin gene of Serratia marcescens. The Ind gene was detected in 21.8% of the S. Typhi isolates from the East Indonesian archipelago, all of which contained the Hd gene. The Hj gene was not detected. The z66 gene was present in 15.4% of the isolates. The presence of these “foreign” flagellin genes could be associated with an increased risk for developing severe disease.

Distribution Flagellin Gene Variants of Salmonella Typhi in Patients with Typhoid Fever in West Kutai, East Kalimantan, Indonesia

“…Detection of flagellin gene encoding Z66 gene using a primer z66_F 5'ATG CAA GCA GTC AATAC ATC-3 'and z66_R 5'TTAACGCAGCAGAGACAGTAC-3' which will generate 1500 bp fragment. Detection of z66Ind using Ind-F: 5 'ATG TCG GAA ATC AAC CGT ATC T 3' and Ind-R: 5 'CAG GCC GTC AAC CTG AGA C 3' that will produce fragments 597 bp [11,13]. Results of detection of S. typhi flagellin gene showed in figure 1 and 2 below.…”

“…For z66Ind primer set designated Ind -F: 5' ATG TCG GAA ATC AAC CGT ATC T 3' and Ind-R:5' CAG GCC GTC AAC CTG AGA C 3' were selected for the specific amplification of a 597bp segment of the Ind gene. The PZ66-A and PZ66-B primers are located in the central region of the z66 gene that is largely deleted in the Ind gene and the primers Ind-F and Ind-R are located in the 5' and 3' portion of the Ind gene that shows homology with the z66 gene, but these primers are chosen such that the number of mismatches with this gene is too high to warrant efficient amplification of the z66 gene [11,12]. The Ind-specific PCR was performed with after an initial denaturation at 94°C for 2 min, for 35 cycles at 94°C for 30s, 55°C for 30s, and 72°C for 1 min, followed by an extension step of 72°C for 5 min [11].…”

Relationships between Flagellin Genes Variants of <i>Salmonella</i> <i>enterica</i> Serovar Typhi and Severity of Illness from Acute and Carriers State of Typhoid Fever

“…The Ind-specific PCR was performed with after an initial denaturation at 94°C for 2 min, for 35 cycles at 94°C for 30 s, 55°C for 30 s, and 72°C for 1 min, followed by an extension step of 72°C for 5 min. [1].…”

“…The PZ66-A and PZ66-B primers are located in the central region of the z66 gene that is largely deleted in the Ind gene and the primers Ind-F and Ind-R are located in the 5' and 3' portion of the Ind gene that shows homology with the z66 gene, but these primers are chosen such that the number of mismatches with this gene is too high to warrant efficient amplification of the z66 gene [1,15]. The Ind-specific PCR was performed with after an initial denaturation at 94°C for 2 min, for 35 cycles at 94°C for 30 s, 55°C for 30 s, and 72°C for 1 min, followed by an extension step of 72°C for 5 min.…”

“…The disease causes much morbidity and mortality in developing countries and is characterized by prolonged fever, bacterial growth in cells of the reticuloendothelial system, and significant inflammation of the lymphoid organs of the small intestine. Symptoms of typhoid fever are characterized by fever (90-100%), coated tongue (96.3%), malaise (93.8%), headache (83.8%), nausea (27.5%), constipation (12.5%), abdominal distention (12.5%), diarrhea (11.3%), and melena (8.8%) [1]. Recognition of these symptoms important in preventing the delayed diagnosis of typhoid fever that can lead to severe typhoid infections such as such as extra-intestinal infectious on central nervous system, cardiovascular system, pulmonary system, bone, joints, hepatobiliary system, and genitourinary system [2].…”