The Potential of grxB Gene for Detection of C. sakazakii in Infant Formula Milk Using Real-Time Polymerase Chain Reaction

Cronobacter sakazakii is one of the bacteria that causes food poisoning that contaminates infant formula. This pathogen causes necrotizing enterocolitis, sepsis, and meningitis in infants or neonates with reported case fatality rates ranging from 40% to 80%. Therefore, it is necessary to develop fast and accurate detection of C. sakazakii in infant formula milk. This research aims to develop a method for detecting C. sakazakii bacteria using real-time PCR with high sensitivity, specificity, and accuracy. A rapid detection method using real-time PCR with the target gene grxB successfully detects the presence of C. sakazakii DNA in artificially contaminated formula milk. The results of the real-time PCR test showed that C. sakazakii DNA with a concentration of 53 ng/µL could be amplified by the grxB gene primer pair with a Ct value of 12 and a Tm value of 85.8ºC. The specificity test showed that the grxB primer could differentiate between target and some non-target bacteria. The sensitivity test showed the ability of the grxB primer to detect the smallest concentration of 3,392 pg/µL with a Ct of 24,06. Based on the results obtained, it can be concluded that the grxB primer has the potential to be used as rapid detection method for C. sakazakii bacteria in infant formula using real-time PCR

Validation of the Detection Kit for Pathogenic Bacteria Salmonella typhi Causes Food Poisoning with Real Time Polymerase Chain Reaction

Salmonella typhi is a gram-negative bacteria that causes food poisoning. Salmonella typhi bacteria can cause typhus, which can produce lipopolysaccharide complex endotoxin, which plays an important role in the pathogenesis of typhoid fever. Cases of food poisoning are still common and are one of the causes of death and death in Indonesia. In 2019, there were 77 cases of food poisoning in Indonesia. A fast, accurate, and specific detection method is needed to detect poisoning. In previous studies, the optimum conditions and formulas for detecting these bacteria have been obtained using Real Time PCR. The results of previous studies are used as the basis for the development of a prototype detection kit. In this study, validation was carried out, which aimed to confirm the results of previous studies so that a reproducible and accurate product for the detection of Salmonella typhi bacteria could be obtained. The results of this study showed that fim-C primers for Salmonella typhi were amplified at 95 base pairs (bp) with an annealing temperature of 60ºC and a standard DNA concentration of 50 ng/µL. The results of the Real Time PCR confirmation test of Salmonella typhi bacteria at a pre-denaturation of 3 minutes with a concentration of 10 pmol obtained the Ct value according to thestandard with previous studies. The Ct value obtained was 13.96 for S. typhi bacteria. Based on the results obtained, it can be conclude that the condition validation stage for pure cultures was successfully carried out by producing consistentand reproducible data

Validation of the Detection Kit for Pathogenic Bacteria Salmonella typhi Causes Food Poisoning with Real Time Polymerase Chain Reaction

Salmonella typhi is a gram-negative bacteria that causes food poisoning. Salmonella typhi bacteria can cause typhus, which can produce lipopolysaccharide complex endotoxin, which plays an important role in the pathogenesis oftyphoid fever. Cases of food poisoning are still common and are one of the causes of death and death in Indonesia. In 2019, there were 77 cases of food poisoning in Indonesia. A fast, accurate, and specific detection method is needed to detect poisoning. In previous studies, the optimum conditions and formulas for detecting these bacteria have been obtained using Real Time PCR. The results of previous studies are used as the basis for the development of a prototypedetection kit. In this study, validation was carried out, which aimed to confirm the results of previous studies so that a reproducible and accurate product for the detection of Salmonella typhi bacteria could be obtained. The results of this study showed that fim-C primers for Salmonella typhi were amplified at 95 base pairs (bp) with an annealing temperature of 60ºC and a standard DNA concentration of 50 ng/µL. The results of the Real Time PCR confirmation test of Salmonellatyphi bacteria at a pre-denaturation of 3 minutes with a concentration of 10 pmol obtained the Ct value according to thestandard with previous studies. The Ct value obtained was 13.96 for S. typhi bacteria. Based on the results obtained, it can be conclude that the condition validation stage for pure cultures was successfully carried out by producing consistentand reproducible data

APLIKASI miRNA SEBAGAI BIOMARKER IDENTIFIKASI PADA INVESTIGASI FORENSIK

Mikro RNA (miRNA) adalah molekul RNA non-coding yang mengandung 18-24 nukleotida yang sangat terkonservasi dan terlibat dalam pengaturan banyak mekanisme biokimia dalam tubuh manusia. Beberapa penelitian menunjukkan bahwa miRNA dapat bertindak sebagai penanda dalam beragam identifikasi forensik dalam mengidentifikasi penentuan waktu kematian, cairan tubuh, vitalitas luka, dan bidang forensik lainnya. miRNA dalam cairan tubuh dan jaringan diketahui meningkat akibat patofisiologi yang berubah. Studi tentang miRNA pada bidang forensik menjadi hal menarik ditelusuri karena stabilitas dan spesifisitasnya sehingga dapat menjawab secara definitif informasi penting yang diperlukan untuk penyelidikan dan penuntutan, seperti asal cairan sampel yang diperiksa. Karakteristik miRNA sebagai penanda molekuler forensik selaras dengan ketahanannya terhadap degradasi sehingga cocok sebagai penandaendogen. Namun, kurangnya protokol dalam pengujian forensik secara ilmiah dan rendahnya studi pada penanda molekuler ini dalam biologi forensik memerlukan analisis literatur ilmiah mengenai penggunaan forensik miRNA

Str Locus Mutations in Paternity Case

DNA analysis is widely applied in solving forensic cases, especially Short Tandem Repat (STR) because of its advantages. In identifying the individual, the National Police compared the individual's DNA with that of his parents. Each anal has a pair of DNA fragments of which half are inherited by the father and the remainder by the mother according to Mendel's Law of Segregation. In this study, we compared DNA typing between the child and the mother with the help of PCR extracted by the Chelex method to find the mother fragment and obtain the father fragment. A child is the biological child of the alleged father if he or she has less than 2 exclusion STR loci. The results of this study revealed that all paternal fragments from the child were identical to the DNA fragments of the alleged father, except for one locus, namely CSF1PO which had a mutation. Mutations in the STR locus lower the paternity index, although it can still be concluded that the child is the biological child of the alleged father.   Keywords: Paternity Test, DNA, STR, Mutatio

The Potential of

Cronobacter sakazakii is one of the bacteria that causes food poisoning that contaminates infant formula. This pathogen causes necrotizing enterocolitis, sepsis, and meningitis in infants or neonates with reported case fatality rates ranging from 40% to 80%. Therefore, it is necessary to develop fast and accurate detection of C. sakazakii in infant formula milk. This research aims to develop a method for detecting C. sakazakii bacteria using real-time PCR with high sensitivity, specificity, and accuracy. A rapid detection method using real-time PCR with the target gene grxB successfully detects the presence of C. sakazakii DNA in artificially contaminated formula milk. The results of the real-time PCR test showed that C. sakazakii DNA with a concentration of 53 ng/µL could be amplified by the grxB gene primer pair with a Ct value of 12 and a Tm value of 85.8ºC. The specificity test showed that the grxB primer could differentiate between target and some non-target bacteria. The sensitivity test showed the ability of the grxB primer to detect the smallest concentration of 3,392 pg/µL with a Ct of 24,06. Based on the results obtained, it can be concluded that the grxB primer has the potential to be used as rapid detection method for C. sakazakii bacteria in infant formula using real-time PCR

Validation of the Detection Kit for Pathogenic Bacteria

Salmonella typhi is a gram-negative bacteria that causes food poisoning. Salmonella typhi bacteria can cause typhus, which can produce lipopolysaccharide complex endotoxin, which plays an important role in the pathogenesis oftyphoid fever. Cases of food poisoning are still common and are one of the causes of death and death in Indonesia. In 2019, there were 77 cases of food poisoning in Indonesia. A fast, accurate, and specific detection method is needed to detect poisoning. In previous studies, the optimum conditions and formulas for detecting these bacteria have been obtained using Real Time PCR. The results of previous studies are used as the basis for the development of a prototypedetection kit. In this study, validation was carried out, which aimed to confirm the results of previous studies so that a reproducible and accurate product for the detection of Salmonella typhi bacteria could be obtained. The results of this study showed that fim-C primers for Salmonella typhi were amplified at 95 base pairs (bp) with an annealing temperature of 60ºC and a standard DNA concentration of 50 ng/µL. The results of the Real Time PCR confirmation test of Salmonellatyphi bacteria at a pre-denaturation of 3 minutes with a concentration of 10 pmol obtained the Ct value according to thestandard with previous studies. The Ct value obtained was 13.96 for S. typhi bacteria. Based on the results obtained, it can be conclude that the condition validation stage for pure cultures was successfully carried out by producing consistentand reproducible data

Validation of the Detection Kit for Pathogenic Bacteria

Salmonella typhi is a gram-negative bacteria that causes food poisoning. Salmonella typhi bacteria can cause typhus, which can produce lipopolysaccharide complex endotoxin, which plays an important role in the pathogenesis of typhoid fever. Cases of food poisoning are still common and are one of the causes of death and death in Indonesia. In 2019, there were 77 cases of food poisoning in Indonesia. A fast, accurate, and specific detection method is needed to detect poisoning. In previous studies, the optimum conditions and formulas for detecting these bacteria have been obtained using Real Time PCR. The results of previous studies are used as the basis for the development of a prototype detection kit. In this study, validation was carried out, which aimed to confirm the results of previous studies so that a reproducible and accurate product for the detection of Salmonella typhi bacteria could be obtained. The results of this study showed that fim-C primers for Salmonella typhi were amplified at 95 base pairs (bp) with an annealing temperature of 60ºC and a standard DNA concentration of 50 ng/µL. The results of the Real Time PCR confirmation test of Salmonella typhi bacteria at a pre-denaturation of 3 minutes with a concentration of 10 pmol obtained the Ct value according to thestandard with previous studies. The Ct value obtained was 13.96 for S. typhi bacteria. Based on the results obtained, it can be conclude that the condition validation stage for pure cultures was successfully carried out by producing consistentand reproducible data