BACKGROUND AND OBJECTIVE

Nontuberculous mycobacteria (NTM) lung disease appears like tuberculosis infection but is resistant to primary anti-tuberculosis drugs. Hence, patients whose sputum sample tests positive for acid-fast bacilli (AFB) and bacterial culture for several times should be assessed for colonization or infection with NTM in a damaged lung secondary to TB. In such cases, though drug-resistant TB may be adequately treated, treatment may need to be directed towards the NTM as well. In NTM therapy, the duration and choice of treatment agent is based upon the specific organism and disease extent. This study used one-step multiplex PCR (mPCR) assay for rapid differentiation of solid cultures in Ogawa medium as Mycobacterium tuberculosis (MTB) and/or NTM.

A total of 80 stocked isolates obtained from the Lung Center of the Philippines from January to December 2018 were screened for NTM in terms of growth in Ogawa medium, acid fastness, and MPT64 TB antigen test result. These were from sputum specimens of multidrug-resistant tuberculosis (MDR-TB) patients. DNA was extracted from cultures (n=55) grown in Ogawa medium and one-step mPCR was performed to identify NTM to the species level.

Out of 80 samples screened, a total of 55 isolates were identified as NTM. One-step mPCR identified 12.73% (7/55) as M. abscessus, 34.55% (19/55) as M. massiliense, 1.82% (1/55) as M. kansasii, and 50.91% (28/55) were identified only up to genus Mycobacteria spp. Neither M. avium complex nor M. intracellulare was identified among the samples tested.

One-step mPCR was able to identify isolates as MTB or NTM coinciding with the initial screening using MPT64 TB antigen test. Multiplex PCR has given a more specific identificati on to the species level. The use of mPCR in identifying MTB and clinically significant NTM’s is suitable for the adequate treatment of mycobacterial infection.

Background: The multiplex polymerase chain reaction can rapidly differentiate taenia spp adults and cysticercose cysts.\r\n', u'Objective: The study aimed to identify taenia spp adults and cysticercose cysts in human by using multiplex PCR\r\n', u'Subjects and methods: The multiplex PCR was applied by using the 4 forward and one reverse primer to amplify the target gene cytochrome c oxydase subunit I (COXI) of the Taenia spp in human. T\r\n', u'Results:4 molecular sizes of PCR products were appeared: 269 bp,720 bp, 827 bp, 984 bp. Seventy six samples including 65 flat worms and 11 cysticercose cysts which collected from the patients who are living in 19 different provinces and cities of Northern part of Viet Nam and treated in the clinic of NIMPE were examined. Of 65 flat worms analyzed 35 samples were T.asiatica (58.46%), 27 were T.saginata (41.54%). All 11 cysticercose cysts were T.solium (100%). \r\n', u'Conclusion: The result also indicated that some time 3 Taenia species were found at the same area. A remarkable difference of infection rate was found between men and women, also adults and children. \r\n', u'\r\n', u'
Taenia spp adults ; cysticercose cysts ; multiplex PCR

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

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

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