scholarly journals KARAKTERISASI DAN IDENTIFIKASI MOLEKULER BAKTERI PENGHASIL ENZIM PROTEASE DARI TEMPE YANG DIPERJUALBELIKAN DI PASAR LUBUK PAKAM

2021 ◽  
Vol 4 (1) ◽  
pp. 7-13
Author(s):  
Visensius Krisdianilo ◽  
Sa'adah Siregar ◽  
Vincentia Ade Rizky
Keyword(s):  
Food Products ◽  
Soybean Seeds ◽  
Protease Enzyme ◽  
Gene 16S Rrna ◽  
Long Time ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Positive Results ◽  
Blast Program

Protease enzyme is an enzyme that is important in protein breakdown. Animals, plants as well as microorganisms such as bacteria can produce this protease enzyme. In its application protease enzymes can be used in the pharmaceutical industry, detergent industry, skin products as well as food products. Tempe is one of the traditional food products that have been known for a long time, tempeh is made from soybean seeds fermented by mushrooms. Molecular identification can use polymerase chain reaction (PCR) method, PCR is the process of multiplying a certain nucleotide sequence using enzymatic processes in vitro. The presence of protein content in tempeh can be possible the presence of bacteria that can break down proteins in the tempeh, especially tempeh that has been fermented about 48-72 hours. Based on the results of characterization and identification of 5 isolates of tempeh post-fermentation 72 hours, positive results of protease enzymes found in isolate TPLP-1, TPLP-2 and TPLP-5, with the largest zone diameter in isolate TPLP-2 50 mm, then isolate with the highest protease enzyme activity isolate TPLP-2 molecularly identified by identifying the gene 16S rRNA which is subsequently included in the BLAST program and obtained by isolate TPLP-2 identified as Pseudomons stuastzeri.

Prevalence of SHV-extended spectrum β-lactamase producing carbapenem –resistantKlebsiellapneumoniae among patients with lower respiratory tractinfections in Babylon Province-Iraq.

2018 ◽  
Vol 9 (SPL1) ◽  
Author(s):  
Fatima Moeen Abbas
Keyword(s):  
Resistance Rate ◽  
Combination Method ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Carbapenem Resistant ◽  
Extended Spectrum ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Tract Infections ◽  
Positive Results

This study was carried out to screen the prevalence of Klebsiella pneumoniae isolated from patients with lower respiratory tract infections in Babylon province.From December,2015 to the end of March,2016,a total of 100 sputum samples were collected from patients visited or hospitalized Merjan Teaching Hospital and Al- Hashimya General Hospital. Fifteenth (65%) isolates were identified as Klebsiellapneumoniae. All bacterial isolates were evaluated for extended spectrum β-lactamase (ESBL) production phenotypically using disk combination method. Eleven (73.3%) isolates were detected as ESBL-producers. Kirby-Bauer disk diffusion method was employed to determine resistance profile of ESBLs-positive isolates. Higher rates of resistance were observed for ampicillin and piperacillin antibiotics with (81.8%) and (72.7%) resistance rate, respectively, while the lowest rate was noticed for imipenem antibiotic (14.28%). Carbapenem-resistant isolates were investigated for blaSHV gene by Polymerase Chain Reaction (PCR) method, 2 (100%) isolates gave positive results.

Identification of Bacteroides forsythus in Subgingival Dental Plaque with the Aid of a Rapid PCR Method

1997 ◽  
Vol 76 (7) ◽  
pp. 1376-1380 ◽  
Author(s):  
J.H. Meurman ◽  
J. Wahlfors ◽  
A. Korhonen ◽  
P. Alakuijala ◽  
P. Väisänen ◽  
...  
Keyword(s):  
Dental Plaque ◽  
Subgingival Plaque ◽  
Chain Reaction ◽  
Rapid Pcr ◽  
Microbiological Methods ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Pre Treatment ◽  
Culture Positive ◽  
Positive Results

Bacteroides forsythus has been shown to be prevalent among patients with periodontitis. Conventional microbiological methods used to identify this bacterium, however, are laborious and time-consuming and are therefore not well-suited for screening purposes. We have developed a polymerase chain-reaction (PCR) method which is rapid, specific, and simple to perform and does not require other sample pre-treatment except a brief centrifugation. This method was applied to the detection of B. forsythus in subgingival plaque of 58 periodontitis patients. When compared with the results of conventional culturing, the PCR method always confirmed the culture-positive results, while none of the PCR negative samples was shown to be culture-positive. The PCR method appeared to give more than double the number of samples positive for B. forsythus than culturing (89.7% vs. 37.9%). The analysis requires less than 4 hrs to perform, and is specific only to B. forsythus and sensitive enough to detect fewer than 5 bacteria.

scholarly journals Development of multiplex real-time RT-PCR assay for the detection of SARS-CoV-2

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250942
Author(s):  
Huseyin Tombuloglu ◽  
Hussein Sabit ◽  
Ebtesam Al-Suhaimi ◽  
Reem Al Jindan ◽  
Khaled R. Alkharsah
Keyword(s):  
Real Time ◽  
Single Gene ◽  
False Negative ◽  
Diagnostic System ◽  
Virus Detection ◽  
Current Method ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Positive Results ◽  
Good Agreement

The outbreak of the new human coronavirus SARS-CoV-2 (also known as 2019-nCoV) continues to increase globally. The real-time reverse transcription polymerase chain reaction (rRT-PCR) is the most used technique in virus detection. However, possible false-negative and false-positive results produce misleading consequences, making it necessary to improve existing methods. Here, we developed a multiplex rRT-PCR diagnostic method, which targets two viral genes (RdRP and E) and one human gene (RP) simultaneously. The reaction was tested by using pseudoviral RNA and human target mRNA sequences as a template. Also, the protocol was validated by using 14 clinical SARS-CoV-2 positive samples. The results are in good agreement with the CDC authorized Cepheid`s Xpert® Xpress SARS-CoV-2 diagnostic system (100%). Unlike single gene targeting strategies, the current method provides the amplification of two viral regions in the same PCR reaction. Therefore, an accurate SARS-CoV-2 diagnostic assay was provided, which allows testing of 91 samples in 96-well plates in per run. Thanks to this strategy, fast, reliable, and easy-to-use rRT-PCR method is obtained to diagnose SARS-CoV-2.

scholarly journals Comparison of Four Polymerase Chain Reaction Methods for the Rapid Detection of Human Fecal Pollution in Marine and Inland Waters

2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
Dave S. Bachoon ◽  
Cortney M. Miller ◽  
Christen P. Green ◽  
Ernesto Otero
Keyword(s):  
Puerto Rico ◽  
Environmental Samples ◽  
Fecal Pollution ◽  
Chain Reaction ◽  
Bifidobacterium Adolescentis ◽  
Fecal Samples ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Positive Results ◽  
Human Specific

We compared the effectiveness of three PCR protocols for the detection ofBifidobacterium adolescentisand one PCR protocol for detectingBacteroidalesas indicators of human fecal pollution in environmental samples. Quantitative PCR indicated that a higher concentration ofB. adolescentisDNA was recovered from sewage samples on the 0.2 μm filters compared to the 0.45 μm filters, and there was no evidence of qPCR inhibitors in the DNA extracts. With the Matsuki method (1999),B. adolescentiswas detected only in undiluted sewage samples. The King method (2007) performed well and detectedB. adolescentisin all of the sewage dilutions (from undiluted to10−4). In contrast, the Bonjoch approach (2004) was effective at detectingB. adolescentisat lower dilutions (10−3) of sewage samples and it gave false positive results with some (3/8) pig fecal samples. Human-specificBacteroidales(HuBacs) were detected in the lower diluents of sewage samples but was positive in pig (6/8) and cattle fecal samples. PCR detection ofB. adolescentisin marine samples from Puerto Rico and freshwater samples from Georgia indicated that the PCR method of King et al. (2007) and the modified Layton method for HuBac were in agreement in detecting human fecal pollution in most sites.

scholarly journals The PI3KCA and AKT Inhibitory Activities of Litsea Cubeba Lour. Fruits and Heartwoods Towards Hela Cells

2019 ◽  
Vol 7 (9) ◽  
pp. 1422-1424
Author(s):  
Aminah Dalimunthe ◽  
Poppy Anjelisa Zaitun Hasibuan ◽  
Denny Satria
Keyword(s):  
Gene Expression ◽  
Cervical Cancer ◽  
Cell Culture ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Litsea Cubeba ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Inhibitory Activities

AIM: To investigated the activities of chloroform fractions at pH 7 of Litsea cubeba Lour. Fruits and heartwoods (CF-7F and CF-7H) in decrease expression of PI3KCA, Akt-1 and Akt-2 genes towards cervical cancer cell culture (HeLa) experiments in vitro. MATERIAL AND METHODS: CF-7F and CF-7H (12.5 and 25 µg/mL) were tested for its potential inhibition on gene expression of PI3KCA, Akt-1 and Akt-2 genes by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) method. RESULT: CF-7F and CF-7H were showed the activity to reduce the expression of PI3KCA, Akt-1 and Akt-2 genes. CONCLUSION: Our results suggest that CF-7F and CF-7H significantly inhibit the expression of PI3KCA, Akt-1 and Akt-2 genes.

Expression of IGF2 and IGF receptor mRNA in bovine nuclear transferred embryos

Zygote ◽  
2003 ◽  
Vol 11 (3) ◽  
pp. 245-252 ◽  
Author(s):  
Dong-Wook Han ◽  
Sang-Jin Song ◽  
Sang Jun Uhum ◽  
Jeong-Tae Do ◽  
Nam-Hyung Kim ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Donor Cell ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Method ◽  
Igf Receptor ◽  
Polymerase Chain ◽  
Low Efficiency ◽  
Insight Into

Incomplete reprogramming of the donor cell nucleus after nuclear transfer (NT) probably leads to the abnormal expression of developmentally important genes. This may be responsible for the low efficiency of cloned animal production. Insulin-like growth factor 2 (IGF2) and IGF2 receptor (IGF2R) are imprinted genes that play important roles in preimplantation development. To obtain an insight into abnormal gene expression after nuclear transfer, we assessed the transcription patterns of IGF2-IGF2R in single in vitro fertilised and cloned embryos by reverse-transcription polymerase chain reaction (RT-PCR). IGF2R expression did not differ significantly but IGF2 was more highly expressed in cloned embryos than in IVF embryos (p < 0.05). This was confirmed by a quantitative RT-PCR method. Thus, incomplete reprogramming may induce abnormal transcription of IGF2 in cloned embryos.

Evaluation of self-collected urine dip swab method for detection of Chlamydia trachomatis

Sexual Health ◽  
2009 ◽  
Vol 6 (3) ◽  
pp. 213 ◽  
Author(s):  
Anna-Maria Costa ◽  
Christopher K. Fairley ◽  
Suzanne M. Garland ◽  
Sepehr N. Tabrizi
Keyword(s):  
Chlamydia Trachomatis ◽  
Room Temperature ◽  
Reaction System ◽  
Urine Samples ◽  
Postal Service ◽  
Positive Urine ◽  
Polymerase Chain ◽  
Inhibition Control ◽  
Positive Results

Background: The present study is an evaluation of a self-collected urine dip (SCUD) swab as an alternative sampling method for the detection of Chlamydia trachomatis (CT) in urine samples that conforms to postal service regulations in Australia. Methods: Sixty urine samples, previously identified as CT positive were used to prepare SCUD swabs in vitro. In addition, replicate SCUD swabs were prepared from known CT positive urine samples and stored at room temperature, or sent through the postal system. All samples were tested for CT and an inhibition control using the Roche TaqMan 48 Real-time polymerase chain reaction system. Results: Overall, 58/60 (97%) SCUD swabs generated positive CT results. Triplicate SCUD swabs prepared from five known positive urine samples and stored up to 7 days at room temperature, showed positive results in all samples. Ten replicates of SCUD swabs from five known CT positive samples were also tested after being posted from different regions in Australia, with a transit time of 2–7 days, back to the Melbourne laboratory. There was 94% positivity of the SCUD swab samples. Conclusion: The present study demonstrated SCUD swabs to be a sensitive and robust method of self-collecting samples for detection of CT subsequent to sending the samples through the postal service.

scholarly journals The frequency of detection of non-polio enteroviruses in foul and fecal waste waters, water and some food products

2019 ◽  
Vol 95 (6) ◽  
pp. 525-528
Author(s):  
Viktor I. Sergevnin ◽  
M. A. Tryasolobova ◽  
E. V. Kudrevatykh ◽  
E. Zh. Kuzovnikova
Keyword(s):  
Polymerase Chain Reaction ◽  
Surface Water ◽  
Water Samples ◽  
Food Products ◽  
Waste Waters ◽  
Chain Reaction ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Wastewater Samples ◽  
Fecal Waste

In the Perm Territory from 2010 to 2014 155 samples offoul andfecal waste waters, 293 samples of surface water, 827 samples of supply net water, and 57 vegetable and fruit water-washes were examined for the RNA enterovirus agent with the use of polymerase chain reaction (PCR) method. In parallel 155 wastewater samples, 20 samples of surface water, and 4 samples of supply net water were examined for non-polio enterovirus agent with the use of virological methods. In the samples of foul waste waters the RNA enterovirus agent was detected in 74.8 ± 3.4%, and nonpolio enterovirus agent - in 65.1 ± 3.8%. In the samples of surface water the RNA enterovirus agent was detected in 2.3 ± 0.8%; in the area offoul and fecal waste waters the non-polio enterovirus agent was detected in 20.0 ± 4.4% in the process of virological investigation of RNA-positive water samples. In supply net water the RNA enterovirus agent was detected in 0.8 ± 0.3 %, on the surface of vegetables, fruits, and grapes - in 10.5 ± 3.9 %.

scholarly journals IDENTIFIKASI BAKTERI ESCHERICHIA COLI O157:H7 PADA FESES PENDERITA DIARE DENGAN METODE KULTUR DAN PCR

2021 ◽  
Vol 3 (2) ◽  
pp. 118-123
Author(s):  
Vincentia Ade Rizky ◽  
Sa’adah Siregar ◽  
Visensius Krisdianilo ◽  
Asvia Rahayu ◽  
Suventi Syafrina Ginting ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Conventional Method ◽  
Diarrheal Disease ◽  
Culture Method ◽  
Escherichia Coli O157 ◽  
Foodborne Disease ◽  
Laboratory Examination ◽  
Long Time ◽  
Pcr Method ◽  
Positive Results

Escherichia coli O157: H7 is the main cause of foodborne disease in several countries, one of which is diarrhea. Diarrheal disease is still a major problem in Indonesia that needs treatment and study from various aspects. The conventional method of laboratory examination such as culture is a method that is often carried out, but in making the diagnosis requires a long time, the number of samples is large, and the results are less accurate because contamination can occur. Another more accurate technique for detecting Escherichia coli O157: H7 is the PCR technique. This study aims to identify the Escherichia coli O157: H7 bacteria by culture method and PCR. The results showed that the culture method and PCR of 8 isolated samples 4 showed positive results for the bacterium Escherichia coli O157: H7. However, the PCR method is more selective and faster than the culture method.

Characterization of the Original Christmas Disease Mutation (Cysteine 206 → Serine): From Clinical Recognition to Molecular Pathogenesis

1992 ◽  
Vol 67 (01) ◽  
pp. 063-065 ◽  
Author(s):  
Sherryl A M Taylor ◽  
Jacalyn Duffin ◽  
Cherie Cameron ◽  
Jerome Teitel ◽  
Bernadette Garvey ◽  
...  
Keyword(s):  
Nucleotide Substitution ◽  
Novel Mutation ◽  
Factor Ix ◽  
Causative Mutation ◽  
Coding Region ◽  
Body Of Knowledge ◽  
Polymerase Chain ◽  
Splice Junctions

SummaryChristmas disease was first reported as a distinct clinical entity in two manuscripts published in 1952 (1, 2). The eponym associated with this disorder, is the surname of the first patient examined in detail and reported by Biggs and colleagues in a paper describing the clinical and laboratory features of seven affected individuals (3). This patient has severe factor IX coagulant deficiency (less than 0.01 units/ml) and no detectable circulating factor IX antigen (less than 0.01 units/ml). Coding sequence and splice junctions of the factor IX gene from this patient have been amplified in vitro through the polymerase chain reaction (PCR). One nucleotide substitution was identified at nucleotide 30,070 where a guanine was replaced by a cytosine. This mutation alters the amino acid encoded at position 206 in the factor IX protein from cysteine to serine. The non conservative nature of this substitution, the absence of this change in more than 200 previously sequenced factor IX genes and the fact that the remainder of the coding region of this gene was normal, all provide strong circumstantial evidence in favour of this change being the causative mutation in this patient. The molecular characterization of this novel mutation in the index case of Christmas disease, contributes to the rapidly expanding body of knowledge pertaining to Christmas disease pathogenesis.

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