DEVELOPMENT OF A UPL PROBE-BASED REAL-TIME PCR ASSAY OF PORCINE DELTACORONAVIRUS IN TAIWAN

Porcine deltacoronavirus (PDCoV) causes clinical symptoms characterized by severe diarrhea and vomiting in neonatal piglets and pregnant sows, which is similar to those resulted from transmissible gastroenteritis coronavirus (TGEV) and porcine epidemic diarrhea virus (PEDV). Since PEDV was considered as the dominant enteric virus all round the world, PDCoV has been unwittingly overlooked due to its indistinguishable clinical signs with other porcine coronaviruses and relatively low death rates in the pig farm. Specimens which have been previously performed for the detection of PEDV in Animal Disease and Diagnostic Center, National Pingtung University of Science and Technology (NPUST) from January 5, 2015 to January 11, 2016 were examined by a novel universal probe library (UPL) probe-based real-time polymerase chain reaction (PCR). A total of 527 clinical specimens from pigs with diarrhea suspected were examined for PDCoV. Positive rates of PDCoV in small intestine and rectal swab were 4.3% (13/305) and 1.8% (4/222), respectively. Collectively, as to the total specimens, the detection rate is 3.2% (17/527). Our results provide development of a UPL probe-based real-time PCR assay and retrospective investigation of potentially circulating PDCoVs in the field in the whole 2015 and early 2016.

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Detection of Aspergillus species in bone marrow transplant patients

The Journal of Infection in Developing Countries ◽

10.3855/jidc.807 ◽

2010 ◽

Vol 4 (08) ◽

pp. 511-516 ◽

Cited By ~ 12

Author(s):

Parisa Badiee ◽

Abdolvahab Alborzi

Keyword(s):

Bone Marrow ◽

Real Time ◽

Bone Marrow Transplant ◽

Real Time Pcr ◽

Clinical Signs ◽

Marrow Transplant ◽

Aspergillus Species ◽

Pcr Assay ◽

Blood Samples ◽

Transplant Patients

Introduction: Invasive aspergillosis is a severe complication of cytotoxic chemotherapies and bone marrow transplantation (BMT). The aim of this study was to assess the utility of a real-time PCR assay for the early diagnosis of Aspergillus species in blood samples from BMT patients. Methodology: Blood specimens (n = 993) from patients (n = 82) scheduled for BMT were collected prior to transplant and for 100 days post transplantation. The specimens were later tested using an Aspergillus-specific real-time PCR assay. Cultures of clinical samples, along with sonography and computerized tomographic scans, were performed as standard of care. Results: Aspergillus DNA was positive in 94 sequential blood samples from 13 patients with clinical and radiological signs of infection. Samples from three of these patients were PCR-positive for Aspergillus in the first week of admission, prior to transplantation. Four patients with aspergillosis were cured with antifungal agents and nine died. An additional 12 patients without clinical signs of infection were PCR-positive on one occasion each, while two patients with clinical signs of infection were PCR-negative. Compared to routine methods of aspergillosis diagnosis, the respective sensitivity, specificity, negative, and positive predictive values of the PCR method by patient were 86.6%, 82%, 96.5% and 52%. Conclusions: The results show that Aspergillus infections in the blood of bone marrow transplant patients can be dectected by PCR methods. Early detection of Aspergillus infections by PCR has the potential to positively impact patient mortality rate and provide cost savings to hospitals.

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DETECTION OF CANINE PARVOVIRUS TYPE 2 BY A COMMERCIALLY AVAILABLE IN-HOUSE PCR

Taiwan Veterinary Journal ◽

10.1142/s1682648519500069 ◽

2020 ◽

Vol 46 (01) ◽

pp. 37-43

Author(s):

Sheng-Lun Yen ◽

Minh Hoang ◽

Wei-Chen Wang ◽

Ming-Lung Hung ◽

Chi-Hsien Chien ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Canine Parvovirus ◽

Diagnostic Tools ◽

Clinical Samples ◽

Pcr Assay ◽

Diagnostic Center ◽

Low Sensitivity ◽

Canine Parvovirus Type 2

Canine parvovirus type 2 (CPV-2) is an important pathogen that causes anorexia, vomiting, acute gastroenteritis, and bloody diarrhea. CPV-2 was divided into three variants (2a–2c) based on residue 426 of the VP2 protein. Recently, a novel Asian CPV-2c isolate was more prevalent in the Asian continent. The diagnostic tools of CPV-2 are characterized into traditional (such as immunochromatography test) and molecular methods based on their pathogen detection mechanisms. However, the low sensitivity of the immunochromatographic tests is supported by their simplicity and rapid ability to provide results for CPV-2. Surprisingly, some immunochromatography tests showed low sensitivity for the detection of novel CPV-2c compared to a real-time polymerase chain reaction (PCR) assay. Sixty rectal swabs were collected from naturally infected dogs with CPV-2 at the Animal Disease Diagnostic Center, National Pingtung University of Science and Technology, Taiwan, from 2016 to 2019. A novel in-house QubeMDx PCR system was used for diagnosis of three CPV-2 variants and compared to real-time PCR diagnosis. This diagnostic assay detected all of CPV-2 variants within 30 min and the sensitivity was identical to a real-time PCR assay using clinical samples. This study provides evidence that this novel, commercially available in-house PCR assay (QubeMDx PCR system) provides useful, simple, sensitive, reliable, and rapid diagnosis of three CPV-2 variants in veterinary clinics.

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Development of a duplex SYBR GreenⅠ based real-time PCR assay for detection of porcine epidemic diarrhea virus and porcine bocavirus3/4/5

Molecular and Cellular Probes ◽

10.1016/j.mcp.2020.101544 ◽

2020 ◽

Vol 51 ◽

pp. 101544 ◽

Cited By ~ 1

Author(s):

Lan-Lan Zheng ◽

Jian-Tao Cui ◽

Hao-Ying Han ◽

Hua-Lin Hou ◽

Leyi Wang ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Porcine Epidemic Diarrhea Virus ◽

Pcr Assay ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea

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Development of a SYBR Green Multiplex Real Time PCR for Simultaneous Detection of Mycobacterium Tuberculosis and Nocardia Asteroides in Respiratory Samples

Ethiopian Journal of Health Sciences ◽

10.4314/ejhs.v31i2.6 ◽

2021 ◽

Vol 31 (2) ◽

Author(s):

Safar Ali Alizadeh ◽

Amir Javadi ◽

Jalal Mardaneh ◽

Neda Nasirian ◽

Sajjad Alizadeh ◽

...

Keyword(s):

Differential Diagnosis ◽

Mycobacterium Tuberculosis ◽

Real Time ◽

Real Time Pcr ◽

Clinical Symptoms ◽

Simultaneous Detection ◽

Nocardia Asteroides ◽

Pcr Assay ◽

Accurate Treatment ◽

Proper Diagnosis

BACKGROUND፡ Nocardia asteroides and Mycobacterium tuberculosis are worldwide-distributed bacteria. These infectious agents can cause many infections in humans, especially in immunocompromised individuals. Pulmonary infections are more common and have similar clinical symptoms. Proper diagnosis and treatment of these patients are important for accurate treatment and could be lifesaving.METHODS: In this study, a multiplex real-time PCR assay was established for the simultaneous detection of the N. asteroides and M. tuberculosis. Both this homemade multiplex real time PCR and routine commercial tuberculosis tests were performed on 150 pulmonary specimens collected from individuals suspected to have tuberculosis.RESULTS: From 150 specimens, 20 samples were acid fast positive, 14 positives for M. tuberculosis by singleplex real time PCR, 10 positives for N. asteroides by singleplex real time PCR and 2 positives for M. tuberculosis and N. asteroides by multiplex real time PCR whereas 14 samples were positive for M. tuberculosis with commercial test. Differential diagnosis of pulmonary tuberculosis is useful for their proper treatment.CONCLUSION: Our test had good performance for differential diagnosis of tuberculosis and nocardiosis. Therefore, it is recommended to be used to diagnose such patients.

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