Detection of bancroftian filariasis in human blood samples from Sorsogon province, the Philippines by polymerase chain reaction

2001 ◽  
Vol 87 (8) ◽  
pp. 677-679
Author(s):  
Elizabeth Torres ◽  
Bernadette Ramirez ◽  
Ferdinand Salazar ◽  
Ma. Pasay ◽  
Judith Alamares ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Blood ◽  
The Philippines ◽  
Bancroftian Filariasis ◽  
Chain Reaction ◽  
Blood Samples ◽  
Human Blood Samples ◽  
Polymerase Chain

scholarly journals Use of the polymerase chain reaction to detect Toxoplasma gondii in human blood samples.

1992 ◽  
Vol 45 (10) ◽  
pp. 910-913 ◽  
Author(s):  
D O Ho-Yen ◽  
A W Joss ◽  
A H Balfour ◽  
E T Smyth ◽  
D Baird ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Toxoplasma Gondii ◽  
Human Blood ◽  
Chain Reaction ◽  
Blood Samples ◽  
Human Blood Samples ◽  
Polymerase Chain

Evaluation of three polymerase chain reaction techniques for detection ofBrucellaDNA in peripheral human blood

2008 ◽  
Vol 54 (5) ◽  
pp. 352-357 ◽  
Author(s):  
Manal M. Baddour ◽  
Dalal H. Alkhalifa
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Blood ◽  
Target Genes ◽  
Chain Reaction ◽  
Clinical Diagnostic Laboratory ◽  
Gene Encoding ◽  
Diagnostic Laboratory ◽  
Human Blood Samples ◽  
Polymerase Chain ◽  
Pcr Techniques

Brucellosis is a widespread zoonosis. Currently the diagnosis of this zoonosis is based on microbiological and serological laboratory tests. Polymerase chain reaction (PCR) has been used to detect DNA from Brucella . Different target genes, primer pairs, PCR techniques, and extraction procedures have previously been published for Brucella detection. But only a few of these primers have been used in human samples, and only one study has been carried out to compare sensitivity between them. In the present study, 3 sets of primers and 3 different PCR protocols amplifying 3 different regions of the Brucella genome were compared for detection of Brucella DNA in a peripheral-blood PCR assay to conclude which is most suitable for the clinical diagnostic laboratory. These 3 pairs of primers amplify 3 different fragments included in (i) a gene encoding a 31 kDa Brucella abortus antigen (B4/B5), (ii) a sequence 16S rRNA of B. abortus (F4/R2), and (iii) a gene encoding an outer membrane protein (omp-2) (JPF/JPR). Some modifications on the reported techniques were applied during the present work to improve the outcome. The results showed that the B4/B5 primer pair had the highest sensitivity for detection of positive samples (98%), the JPF/JPR primer pair detected 88.4% of positive samples, whereas F4/R2 primer pair was the least sensitive, being able to detect only 53.1% of positive samples. The specificity of the 3 techniques was 100%. The B4/B5 primer pair was also able to detect the smallest number of bacteria (700 cfu/mL), whereas JPF/JPR was able to detect 7 × 105 cfu/mL and F4/R2 was able to detect 7 × 107cfu/mL. It is thus concluded that using the B4/B5 primer PCR with the suggested modifications is a robust assay, which meets the sensitivity requirements to be used for testing of human blood samples for brucellosis in the diagnostic laboratory.

scholarly journals Sporadic Rift Valley Fever Outbreaks in Humans and Animals in Uganda, October 2017–January 2018

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Doreen Birungi ◽  
Freda Loy Aceng ◽  
Lilian Bulage ◽  
Innocent Herbert Nkonwa ◽  
Bernadette Basuta Mirembe ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Blood ◽  
Rift Valley ◽  
Rift Valley Fever ◽  
Suspected Case ◽  
Chain Reaction ◽  
Animal Blood ◽  
Valley Fever ◽  
Blood Samples ◽  
Polymerase Chain

Introduction. Rift Valley fever (RVF) is a mosquito-borne viral zoonosis. The Uganda Ministry of Health received alerts of suspected viral haemorrhagic fever in humans from Kiruhura, Buikwe, Kiboga, and Mityana districts. Laboratory results from Uganda Virus Research Institute indicated that human cases were positive for Rift Valley fever virus (RVFV) by polymerase chain reaction. We investigated to determine the scope of outbreaks, identify exposure factors, and recommend evidence-based control and prevention measures. Methods. A suspected case was defined as a person with acute fever onset, negative malaria test result, and at least two of the following symptoms: headache, muscle or joint pain, bleeding, and any gastroenteritis symptom (nausea, vomiting, abdominal pain, diarrhoea) in a resident of Kiruhura, Buikwe, Mityana, and Kiboga districts from 1st October 2017 to 30th January 2018. A confirmed case was defined as a suspected case with laboratory confirmation by either detection of RVF nucleic acid by reverse-transcriptase polymerase chain reaction (RT-PCR) or demonstration of serum IgM or IgG antibodies by ELISA. Community case finding was conducted in all affected districts. In-depth interviews were conducted with human cases that were infected with RVF who included herdsmen and slaughterers/meat handlers to identify exposure factors for RVF infection. A total of 24 human and 362 animal blood samples were tested. Animal blood samples were purposively collected from farms that had reported stormy abortions in livestock and unexplained death of animals after a short illness (107 cattle, 83 goats, and 43 sheep). Convenient sampling for the wildlife (10 zebras, 1 topi, and 1 impala) was conducted to investigate infection in animals from Kiruhura, Buikwe, Mityana, and Kiboga districts. Human blood was tested for anti-RVFV IgM and IgG and animal blood for anti-RVFV IgG. Environmental assessments were conducted during the outbreaks in all the affected districts. Results. Sporadic RVF outbreaks occurred from mid-October 2017 to mid-January 2018 affecting humans, domestic animals, and wildlife. Human cases were reported from Kiruhura, Buikwe, Kiboga, and Mityana districts. Of the 24 human blood samples tested, anti-RVFV IgG was detected in 7 (29%) human samples; 1 human sample had detectable IgM only, and 6 had both IgM and IgG. Three of the seven confirmed human cases died among humans. Results from testing animal blood samples obtained from Kiruhura district indicated that 44% (64/146) cattle, 46% (35/76) goats, and 45% (9/20) sheep tested positive for RVF. Among wildlife, (1/10) zebras, (1/1) topi, and (1/1) impala tested positive for RVFV by serological tests. One blood sample from sheep in Kiboga district tested RVFV positive. All the human cases were exposed through contact or consumption of meat from infected animals. Conclusion. RVF outbreaks occurred in humans and animals in Kiruhura, Buikwe, Mityana, and Kiboga districts. Human cases were potentially infected through contact with infected animals and their products.

Comparison of Droplet Digital Polymerase Chain Reaction (ddPCR) and Real-Time Quantitative Polymerase Chain Reaction (qPCR) in Detecting Neonatal Invasive Fungal Infections

2021 ◽  
Vol 11 (3) ◽  
pp. 373-379
Author(s):  
Huitao Li ◽  
Xueyu Chen ◽  
Xiaomei Qiu ◽  
Weimin Huang ◽  
Chuanzhong Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Fungal Isolation ◽  
Blood Samples ◽  
Fungal Strains ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Invasive fungal infection (IFI) is the leading cause of death in neonatal patients, yet the diagnosis of IFI remains a major challenge. At present, most IFI laboratory diagnostic methods are based on classical, but limited, methods such as fungal isolation and culture and histopathological examination. Recently, quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) technology have been adopted to quantify nucleic-acid identification. In this study, we established qPCR and ddPCR assays for IFI diagnosis and quantification. qPCR and ddPCR were carried out using identical primers and probe for the amplification of 18S rRNA. Assay results for three fungal strains were positive, whereas ten non-fungal strains had negative results, indicating 100% specificity for both ddPCR and qPCR methods. Genomic DNA of Candida albicans was tested after a serial dilution to compare the sensitivity of the two PCR methods. The limit of detection of ddPCR was 3.2 copies/L, which was a ten-fold increase compared with that of the qPCR method (32 copies/L). Blood samples from 127 patients with high-risk factors and clinical symptoms for IFI were collected from a NICU in Shenzhen, China, and analyzed using qPCR and ddPCR. Thirty-four blood samples from neonates had a proven or probable diagnosis of IFI, and 25 of these were positive by qPCR, whereas 30 were positive by ddPCR. Among the 93 blood samples from neonates who had a possible IFI or no IFI, 24 were positive using qPCR, and 7 were positive using ddPCR. In conclusion, ddPCR is a rapid and accurate pan-fungal detection method and provides a promising prospect for IFI clinical screening.

scholarly journals Evaluation on the presence of Anaplasma, Ehrlichia, and Babesia spp. in goats (Capra hircus) in Cebu, the Philippines

2019 ◽  
Vol 12 (6) ◽  
pp. 774-777 ◽  
Author(s):  
Adrian P. Ybañez ◽  
Orgil V. Arrabis ◽  
Dennis Justin M. Alvarez ◽  
Eloiza May S. Galon ◽  
Rhea Mae P. Jayag ◽  
...  
Keyword(s):  
Blood Smear ◽  
Peripheral Blood Smear ◽  
The Philippines ◽  
Capra Hircus ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Blood Samples ◽  
Blood Smear Examination ◽  
Polymerase Chain ◽  
Babesia Spp

Background: Tick-borne diseases are caused by a wide variety of viruses, pathogens, and diseases. Anaplasma, Ehrlichia, and Babesia spp. are among the most known tick-borne pathogens in Asia. In the Philippines, these pathogens were already reportedly present in dogs and large ruminants, but no study has been reported yet evaluating their presence in goats. Aim: The present study aimed to evaluate the presence of Anaplasma, Ehrlichia, and Babesia spp. in goats in Cebu, the Philippines. Materials and Methods: A total of 100 blood samples from goats were collected in Cebu, the Philippines. Profile of sampled goats including age, body score, and sex was obtained. Peripheral blood smear examination and DNA extraction were performed. Nested polymerase chain reaction assay was used to evaluate the presence of Anaplasma, Ehrlichia, and Babesia spp. Results: None of the samples were found positive with Anaplasma, Ehrlichia, and Babesia spp. infection. Conclusion: Tested goats were negative with Anaplasma, Ehrlichia, and Babesia spp. and calls for continuous surveillance of these pathogens due to the reported detection of these pathogens in other livestock animals in the area.

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