Background: Acute respiratory infection (ARI) is a major cause of morbidity and mortality among children worldwide however, local data on the etiologic diagnosis of ARI are limited. Objectives: To determine the prevalence and the most commonly detected respiratory pathogens using a multiplex PCR assay, known as the Respiratory Panel, among hospitalized children with ARI and compare their clinical and laboratory differences. Methods: This is a cross sectional study of children with ARI who were tested with a multiplex PCR assay. Retrospective chart review was done on these patients admitted from January 2018 to February 2020. Results: There were 47 charts reviewed, mean age was 4.2 years old. Out of 47 patients, 36 (76.6%) tested positive for a pathogen. Respiratory syncytial virus (RSV) being the most common followed by Influenza A/H1-2009 and Human metapneumovirus (hMPV). Two patients had viral co-infections and no bacteria were detected on all subjects. 61.7% patients were started on antibiotics on admission. Fever and cough were the most common sign and symptom, respectively. Normal WBC (68% with neutrophilic predominance) and platelet were detected in 72.3% and 70.2% of patients, respectively; 50% of patients had normal CRP and 60.5% had abnormal findings on chest x-ray. Only the presence of chest x-ray findings was found to have a higher probability of yielding a positive Respiratory Panel p=0.27. Conclusion Among admitted patients with ARI, 76.6% tested positive for a respiratory pathogen. All were caused by viruses presenting as nonspecific manifestations – fever and cough. Clinical manifestations, CBC and CRP showed no association with the Respiratory Panel result while abnormal chest x-ray had a higher probability of yielding a positive Respiratory Panel result.
Multiplex Polymerase Chain Reaction

No abstract available.
Humans* ; Multiplex Polymerase Chain Reaction* ; Papilloma*

Aims: This study was aimed to test the specificity of primers and probes with target genes by using multiplex PCR and multiplex real-time PCR methods. These methods were compared with traditional blood culture methods in detecting five bacteria causing sepsis, including Acinetorbacter baumannii, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus. Methodology and results: A total of 587 blood samples from patients diagnosed with sepsis and septic shock were collected at Thanh Nhan Hospital, Hanoi, Vietnam. Each sample was divided into three parts for bacterial culture, multiplex PCR and multiplex real-time PCR to detect the similarity of the two PCR methods with the bacterial culture method. Conditions in multiplex PCR and multiplex real-time PCR were optimized to ensure the successful amplification of target genes. Results showed that the primers and probes were tested completely specific to the target genes and using multiplex PCR and multiplex real-time PCR techniques could detect five pathogens causing sepsis, including A. baumannii, K. pneumoniae, P. aeruginosa, E. coli and S. aureus. Conclusion, significance and impact of study Both multiplex PCR and multiplex real-time PCR methods have high similarities with the culture method, showing potential in the application of bacteria detection in sepsis.
Multiplex Polymerase Chain Reaction ; Sepsis--microbiology

The aim of this study was to investigate the pattern of distribution of mating type (MAT) genes of Tuber indicum in ectomycorhizosphere soils from natural T. indicum-producing areas and cultivated truffle orchards and ascocarp samples from different regions. Quantitative real-time PCR and multiplex PCR were used to weight the copy numbers of MAT1-1-1 and MAT1-2-1 in natural truffle soils and cultivated orchard soils. The effect of limestone on the pattern of truffle MAT genes and the correlation between soil properties and the proportion of MAT genes were also assessed. These results indicated that an uneven and nonrandom distribution of MAT genes was common in truffle-producing areas, cultivated truffle orchards, and ascocarps gleba. The competition between the two mating type genes and the expansion of unbalanced distribution was found to be closely related to truffle fructification. Limestone treatments failed to alter the proportion of the two mating type genes in the soil. The content of available phosphorus in soil was significantly correlated with the value of MAT1-1-1/MAT1-2-1 in cultivated and natural ectomycorhizosphere soils. The application of real-time quantitative PCR can provide reference for monitoring the dynamic changes of mating type genes in soil. This study investigates the distributional pattern of T. indicum MAT genes in the ectomycorhizosphere soil and ascocarp gleba from different regions, which may provide a foundation for the cultivation of T. indicum.
Calcium Carbonate ; Multiplex Polymerase Chain Reaction ; Phosphorus ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Soil

No Abstract Available.
Korea* ; Multiplex Polymerase Chain Reaction* ; Salmonella enteritidis* ; Salmonella typhimurium* ; Salmonella*

Multiplex Ligation dependent Probe Amplification (MLPA) to detect large deletions or duplications has been widely used as a diagnostic tool for various disease clinically. As this method requires only a small amount of template DNA and is very simple and high throughput, it has numerous advantages for the analysis of the human specimen obtained from archaeological sites. In this study we therefore tried to perform MLPA analysis for detecting any of duplications or deletions in mummy samples (n=4) from medieval Joseon tombs of Korea. Of them, we could not get any authentic data from 3 samples by MLPA method while only one case (HD2) showed the possible presence of duplications or deletions during her lifetime. Although the current report reveal that MLPA is a promising tool for anthropological study in South Korea, more studies are still needed to make up for the validity problem of commercial MLPA kit used in this study.
DNA ; Humans ; Korea* ; Multiplex Polymerase Chain Reaction* ; Mummies

Seahorse is one the most commonly used medicinal animal in China. Five species of Hippocampus are recorded as seahorse in the Chinese Pharmacopoeia. Because of the rapid decrease, several other Hippocampus species are often adulterants as medicinal seahorse in the herbal market, which compromise clinical efficacy and pose threat to endangered seahorse species conversation. Herein, a multiplex polymerase chain reaction (mPCR) method was developed to identify the biological sources of medicinal seahorses.Based on the sequences of mitochondrial DNA, five specific primers for Hippocampus trimaculatus, H. kelloggi, H. kuda, H. histrix and H. mohnikei (H. japonicus)were designed, respectively. Multiplex PCR yields the products of 155, 222, 292, 352, 458 bp amplicons in the present of DNA templates of H. kuda, H. mohnikei, H. kelloggi, H. histrix and H. trimaculatus, respectively. This multiplex PCR method which electrophoresis migration of different lengths of DNA bands allowed simultaneous identification of all the five medicinal seahorses in a single assay. It showed that this multiplex PCR assay is useful for the simultaneous identification the biological sources of complex multi-source samples, which could provide a useful tool for the quality control of seahorses.
Animals ; DNA Primers ; DNA, Mitochondrial ; Multiplex Polymerase Chain Reaction ; Smegmamorpha

Twenty-five isolates of Fusarium fujikuroi acquired from rice seeds and rice plants evidencing symptoms of Bakanae disease were evaluated to determine their mating types and characterize the formation of their sexual state. The mating types of the isolates were evaluated via multiplex PCR with the diagnostic primers of the mating-type (MAT) region: GFmat1a, GFmat1b, GFmat2c, and GFmat2d. Among the 25 isolates, 11 were identified as MAT-1 (male), and 14 as MAT-2 (female). Four MAT-1 isolates and three MAT-2 isolates were mated and cultured to evaluate the optimal culture conditions for the production of their sexual states. Among four tested media, 10% V8 juice agar proved optimal for the perithecial production of the isolates. The isolates also generated the largest numbers of perithecia when incubated at 23degrees C in alternating cycles of 12 hr fluorescent light and NUV fluorescent light and 12 hr darkness.
Agar ; Darkness ; Fusarium ; Light ; Multiplex Polymerase Chain Reaction ; Seeds

To study the correlation of human papillomavirus (HPV) infection with clinical stage in cervical abnormalities, 17 cases of normal cervical tissue and 69 cases of abnormal cervical tissue (cervical intraepithelial neoplasia and invasive cervical cancer) was examined by PCR with HPV-specific consensus primers. One case (5.9%) of normal cervical tissue and 42 cases (60.9%) of abnormal cervical tissues harbored HPV. To investigate the integration of HPV genome in 24 cases of HPV 16 positive cervical cancer, E2 gene of HPV 16 was amplified. Integration of HPV 16 was found in 7 cases (29.2%) with E2 disruption. All samples with E2 disruption were from invasive cervical cancer. A multiplex PCR for the mapping of integrated HPV 16 genome with an anchor primer and indicator primers showed that 11 cases (45.8%) were disrupted somewhere in HPV genome but E6, E7, and LCR regions were conserved in all cases. Seven types of integrated HPV genome from long- (7,062 bp) to short-conserved type (3,204 bp) with various deletions were detected by the multiplex PCR. These results show that integration can be detected more accurately by multiplex PCR than by E2 PCR, and E2 disruption is not a critical event of integration
Consensus ; Genome ; Human papillomavirus 16 ; Humans* ; Multiplex Polymerase Chain Reaction ; Polymerase Chain Reaction ; Uterine Cervical Neoplasms*

Multiplex PCR for three STRs of same repetition unit [GATA]n, 4804LR[D12S66], 27H39LR[DYS19] and 4815LR[D12S67] loci, was constructed for forensic application DNA was extracted from 200 unrelated Koreans and amplified with a mixture of polyacrylamide gel electrophoresis, so called Amp-FLP procedure. Three loci could be co-amplified in a reaction with easy, and reaction condition was not so quite different from that of each locus. The PCR products of each locus could be separated bp, and 4815LR from 241 bp to 281 bp, so these alleles of each locus could be separated on a single electrophoresis. A total of six alleles was noted in 4804LR and heterozygosity was 0.5764. The allele 11 and allele 12 were frequently noted with the frequency of 0.6225 and 0.1775, respectively. Sequencing was done for 2 alleles, and the exact size of the alleles and the repetition unit were confirmed. Through statistical analysis forensic applicability of the STR 4804LR locus was confirmed. For 4815LR and heterozygosity was 0.5764. The allele 11 and llele 12 were frequently noted with the frequency of 0.6225 and 0.1775, respectively. Sequencing was done for 2 alleles, and the exact size of the alleles and the repetition unit were confirmed. Through statistical analysis forensic applicability of the STR 4804LR locus was confirmed. For 4815LR locus the amplification was successful, but the separation of the alleles on routine polyacrylamide gel was not successful. Some alleles was hardly separable, some alleles did not match the allelic ladder exactly, so the interallele was suspicious. On sequencing gel the electrophoresis pattern was quite different with that of routine polyacrylamide gel. A total of 11 allele was noted in 4815LR and heterozygosity was 0.765. For the routine use of the 4815LR locus, more meticulous method for the separation of the alleles such as using automatic DNA sequencer was necessary.
Alleles ; DNA ; Electrophoresis ; Electrophoresis, Polyacrylamide Gel ; Multiplex Polymerase Chain Reaction* ; Polymerase Chain Reaction