Displaying publications 121 - 140 of 1868 in total

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  1. Vongnady K, Rucksaken R, Mangkit B
    Trop Biomed, 2020 Sep 01;37(3):691-712.
    PMID: 33612783 DOI: 10.47665/tb.37.3.691
    Haemonchus spp. or barber's pole worms are one of the most highly pathogenic nematodes of ruminants causing economic losses in livestock worldwide. The current study was a first attempt to identify Haemonchus spp. from goats in Thailand and Lao PDR. Utilizing the inexpensive tools of the discriminant function (DF) combined with synlophe patterns is fundamental for understanding their epidemiological aspects. In total, 255 randomly chosen adult male Haemonchus worms from goats in various areas in each country were identified individually. For both these countries, about 94% based on the DF values, and 99%, 98%, and 97% based on synlophe patterns in the region of the esophageal intestinal junction (EI), 4 mm from the anterior end, and at both these positions, respectively, were identified as H. contortus. Other identified specimens defined as H. placei and hybrids as well as unclassified species based on synlophe patterns were proved using polymerase chain reaction (PCR); this also included some randomly chosen H. contortus by DF and synlophe patterns. All those specimens were confirmed as H. contortus being strongly supported by some genetic evidences and UPGMA analysis. Thus, it was assumed that all specimens in the current study were H. contortus. The morphological differences of this predominant species (H. contortus) in goats between the two countries were: body length, gubernaculum length, and left spicule barb length, while almost all characters of male worms individually measured appeared to overlap, mostly in H. contortus and H. placei, which may lead to misclassification. Therefore, using the DF along with synlophe patterns can assist in increasing the accuracy of Haemonchus spp. identification from goats in some areas where funding is limited, particularly in Lao PDR. The present results revealed that synlophe patterns in the EI region seemed to be promising for the identification of Haemonchus spp., while molecular techniques are also required to address ambiguous identification with some specimens.
    Matched MeSH terms: Polymerase Chain Reaction
  2. Chávez M, Cabezas AF, Ferauds M, Castillo JE, Caicedo LD
    Trop Biomed, 2020 Sep 01;37(3):650-662.
    PMID: 33612779 DOI: 10.47665/tb.37.3.650
    Pseudomonas aeruginosa is considered an opportunistic pathogen, causing a wide variety of infections in compromised hosts, also frequently develops multi-resistance to antibiotics and can colonize various habitats, including water systems. The main aim of this study was to investigate antibiotics susceptibility pattern, genotypic diversity and detection of resistence genes in P. aeruginosa isolates from clinical and aquatic environment sources. Of the 220 P. aeruginosa isolates examined, 48 were clinical isolates and 172 isolates from wastewater and freshwater. Susceptibility to eight antimicrobial agents was carried out by disk diffusion method. Clinical and environmental isolates were screened for the presence of the genes encoding blaKPC-2, blaCTX-M-9, blaPER-1, blaOXA-10, blaIMP-1, blaVIM-2 and blaampC by polymerase chain reaction (PCR). Isolates were examined with PCR-SSCP analysis of partial DNAr 16S sequence. Isolates were mainly resistant to cefoxitin. Multidrug-resistant P. aeruginosa (MDRPA) strains were found in 70% and 90.3% of the clinical and environmental isolates, respectively. The prevalence rates of â-lactamase genes were recorded (blaKPC-2 41.3%, blaVIM-2 36.8%, blaIMP-1 13.6%, blaCTX-M-9 10.9% and blaampC 10.5%,). The PCR-SSCP analysis showed three conformational patterns. All clinical isolates and most environmental isolates were grouped into a single cluster. In this study, we found that P. aeruginosa strains recovered from city water systems must be considered potential reservoir for ESBL genes, especially blaKPC-2 and blaVIM-2.
    Matched MeSH terms: Polymerase Chain Reaction
  3. Nimsuphan B, Prasroedsang S, Kengradomkij C, Thayananuphat A, Kromkhun P
    Trop Biomed, 2020 Sep 01;37(3):551-559.
    PMID: 33612770 DOI: 10.47665/tb.37.3.551
    Uveitis associated with Ehrlichia canis or Anaplasma platys infections were reported in dogs. However, only two E. canis-infected dogs with hypergammaglobulinemia showed acute blindness were reported. There were limited data of the species of Ehrlichia or Anaplasma and the alteration of serum protein fractions in infected dogs. Thus, the species of causative pathogen were investigated and compared the serum protein fractions between infected dogs associated with anterior uveitis and panuveitis in clinical situations. All 103 studied dogs were brought into the ophthalmology clinic which each dog showed signs of unilateral or bilateral uveitis related to ehrlichial infection. Dogs were divided into anterior uveitis and panuveitis groups. The species of Ehrlichia or Anaplasma were identified using nested-PCR based on the 16S rRNA gene and DNA sequencing from blood samples. The serum protein fractions were analyzed using electrophoresis. Fifty-eight dogs (56.31%) were positive of which E. canis and A. platys were detected in 51 and 7 dogs, respectively. The total serum protein and globulin levels were higher in the infected dogs associated with panuveitis than anterior uveitis while the albumin levels were significantly lower in the panuveitis group. The A/G ratios significantly decreased in both groups. Gamma globulin was detected at high levels in both groups while beta globulin significantly increased in the panuveitis group. Hypergammaglobulinemia was detected in 76.92 and 90.90% of infected dogs associated with anterior uveitis and panuveitis, respectively. Most of the infected dogs associated with panuveitis showed significantly levels of hyperproteinemia, hyperbetaglobulinemia and hypergammaglobulinemia compared with anterior uveitis group. E. canis was found as the major pathogen in infected dogs associated with uveitis in this study.
    Matched MeSH terms: Polymerase Chain Reaction
  4. Alshahrani MY, Alanazi AD, Alouffi AS, Abdullah HHAM, Allam AM, Mahmoud MS, et al.
    Trop Biomed, 2020 Sep 01;37(3):587-598.
    PMID: 33612774 DOI: 10.47665/tb.37.3.587
    Knowledge of molecular identification of tick-borne pathogens in camels in Saudi Arabia is very limited; few molecular epidemiological studies have been under taken. This study was to detect Anaplasma spp. and Piroplasma spp. in camels from Asir Province, Saudi Arabia. A total of 150 blood samples were collected from camels in Asir Province and investigated by polymerase chain reaction (PCR) that targeted 18S rRNA and 23S rRNA to detect the DNA of Piroplasma spp. and Anaplasma spp., respectively. The positive samples for 23S rRNA were assayed again by PCR targeting the 16S rRNA. All the blood samples were free from Piroplasma spp. infection. Three camels (2%) were found to be positive for Anaplasma infection through use of PCR that targeted the 23S rRNA gene. There were no significant differences between ages or sexes in the camels that tested positive for Anaplasma. All positive Anaplasma infections were recorded in camels that were infested by ticks. Two Anaplasma sequences for the16S rRNA gene were deposited in GenBank with accession numbers MN882724 and MN882725. They recorded 99.16% and 99.34% similarities (respectively) with KF843825.1 (Candidatus Anaplasma camelii reported in Unizah, Saudi Arabia). Phylogenetic analyses revealed that the two sequences recorded in this study were close to each other; both were located in one cluster with Candidatus Anaplasma camelii isolates that were recorded before in the adjacent areas of Unizah in Saudi Arabia and Iran. In conclusion: two new Anaplasma genotypes close to Candidatus Anaplasma camelii were found in camels in Asir Province, Saudi Arabia for the first time. The camels in this province were found to be free of Piroplasma infection.
    Matched MeSH terms: Polymerase Chain Reaction
  5. Wahedi JA, Ande AT, Oduola AO, Obembe A, Tola M, Oyeniyi TA, et al.
    Trop Biomed, 2020 Sep 01;37(3):637-649.
    PMID: 33612778 DOI: 10.47665/tb.37.3.637
    Studies profiling community and zonal malaria entomological risk indices are required to identify high risk areas where targeted control resources are most needed or likely to have the greatest impact on reducing risk of malaria infection. This study presents a first report on malaria vector risk indices in two vegetation zones within Adamawa state, Nigeria. Endophilic mosquitoes were collected for one year in selected communities in the Guinea and Sudan savanna zones within the State. Plasmodium falciparum Sporozoite and human blood meal ELISA assays were carried out on the female Anopheles mosquitoes collected. Sibling species composition of the An. gambiae complex were determined using PCR assays. Mean numbers of mosquitoes in the Guinea savanna communities were significantly (t = 7.73, DF = 11, p < 0.001) higher than the Sudan. Man-biting rates (F = 2.76, p = 0.13) of Anopheles mosquitoes were higher in the Guinea but not significantly different from Sudan savanna. Sporozoite rates of mosquitoes within the Guinea savanna were 2.7 times higher than the Sudan. The predominant Anopheles coluzzii species encountered in the state had higher overall human blood indices (0.63) and sporozoite rates (6.9%) compared to An. gambiae (0.39, 1.9%) and An. arabiensis (0.58, 2.3%) respectively. Overall annual human blood indices (0.59) of mosquitoes in Adamawa were lower compared to reports from other States. Prevalence and higher transmission risks indices of endophilic An. coluzzii mosquitoes reveal the need for LLIN and management of relatively permanent An. coluzzii breeding sites in the State. Widespread cattle rearing lifestyle and lower human blood indices of mosquitoes in the study area suggest the need to investigate cattle blood indices of the mosquitoes in the state. Higher entomological risk indices in the Guinea Savanna zone provide baseline information for prioritization of malaria vector control supplies within the State.
    Matched MeSH terms: Polymerase Chain Reaction
  6. Sulehria MU, Ahmad SS, Ijaz M, Mushtaq MH, Khan AY, Ghaffar A
    Trop Biomed, 2020 Dec 01;37(4):963-972.
    PMID: 33612749 DOI: 10.47665/tb.37.4.963
    Canine Enteric Coronavirus (CCoV) is one of the major enteric pathogen affecting dogs. This study aims to investigate the molecular prevalence, phylogenetic analysis, associated risk factors, and haemato-biochemical alterations in Canine Coronavirus in dogs in district Lahore, Pakistan. 450 fecal samples were collected from symptomatic dogs originating from various pet-clinics and kennels during 2018-2019. Samples were initially analyzed by sandwich lateral flow immunochromatographic assay and then further processed by RT-PCR (reverse transcriptase polymerase chain reaction) targeting the M gene followed by sequencing. RT-PCR based positive (n=20) and negative (n=20) dogs were samples for their blood for the haemato-biochemical analysis. A questionnaire was used to collect data from pet owners, in order to analyze the data for risk factors analysis by chi square test on SPSS. The prevalence of CCoV was 35.1%, and 23.8 % through Sandwich lateral flow immunochromatographic and RT-PCR respectively. Various risk factors like breed, age, sex, vomiting, diarrhea, sample source, body size, cohabitation with other animals, living environment, food, deworming history, contact with other animals or birds feces, and season were significantly associated with CCoV. The CCoV identified in Pakistan were 98% similar with the isolates from China (KT 192675, 1), South Korea (HM 130573, 1), Brazil (GU 300134, 1), Colombia (MH 717721, 1), United Kingdom (JX 082356, 1) and Tunisia (KX156806). Haematobiochemical alterations in CCoV affected dogs revealed anaemia, leucopenia, lymphopenia, neutrophilia, and decreased packed cell volume, and a significant increase in alkaline phosphate and alanine transaminase. It is concluded that infection with canine coronavirus appears widespread among dog populations in district Lahore, Pakistan. This study is the first report regarding the molecular detection and sequence analysis of CCoV in Pakistan.
    Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction/veterinary
  7. Ashouri Saheli Z, Shenagari M, Harzandi N, Monfared A
    Trop Biomed, 2019 Sep 01;36(3):766-775.
    PMID: 33597498
    The need for an intensive care protocol, sometimes weekly or biweekly, has led to a significant increase in laboratory costs for kidney recipients. In the present study, an inhouse tetraplex nested PCR assay was developed and validated for the specific detection of BKV, JCV, HCMV and EBV in clinical samples. We determined the Limit of Detection (LOD) and analytical specificity. To demonstrate the diagnostic performance of the assay, a total of 102 archival plasma samples were tested and compared with a commercial uniplex real-time PCR kits. The analytical sensitivity of the in-house tetraplex nested PCR assay was 173 copies/ml, when all four viruses were present in the specimens. These values were 79.2, 58.7, 87.6 and 96.1 copies/ml when only, BKV, JCV, HCMV and EBV respectively, were present. The cross-reactivity assays were shown no detectable signal in the tetraplex PCR results. The estimated diagnostic sensitivities were 92.6% for BKV, 92.3% for JCV and 100% for both HCMV and EBV as compared with commercial kits. Regarding the sensitivity and specificity, it seems that the developed Multiplex Nested PCR assay could be used as a reliable virusassociated renal rejection (VRR) panel in post renal transplant surveillance.
    Matched MeSH terms: Real-Time Polymerase Chain Reaction
  8. Kulpraneet M, Limtrakul A, Thanomtham P, Taemaitree N, Puttikamonkul S, Pongsunk S, et al.
    Trop Biomed, 2019 Dec 01;36(4):874-882.
    PMID: 33597460
    Tuberculosis (TB) is a leading cause of morbidity and mortality in Thailand. Cytokines play important roles in defense against Mycobacterium tuberculosis infection. Interleukin (IL)-4 is one of the anti-inflammatory cytokines and has been found to be elevated in TB patients. The common polymorphisms in IL-4 gene, including IL-4-590C/T, IL-4-33C/T, and IL-4-variable number of tandem repeats (VNTR) intron 3 have been reported to be associated with risk for some diseases. The purpose of this study was to investigate possible associations between the above mentioned three common functional polymorphisms in the IL-4 gene in patients with pulmonary tuberculosis (PTB) in a Thai population. Forty three patients with PTB and 90 healthy control subjects were studied. The three common polymorphisms of the IL-4 gene were determined using polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). The allele and genotype frequencies of IL-4 -590 C/T, -33 C/T, VNTR intron 3 polymorphisms did not show significant differences between PTB patients and healthy controls (genotype: p=0.88, p=0.92, p=0.40; allele: p=0.38, p=0.44, p=0.53, respectively). However, the allele distribution of the IL-4 -590 C, -33 C, and VNTR R3 was higher among PTB patients (25.58%, 25.58%, 25.58%, respectively) than among control subjects (20%, 20.48%, 19.44%, respectively). This may suggest that IL-4-590C/T, -33C/T and VNTR intron 3 might play a role in susceptibility to PTB. A larger cohort may possibly help conclude our findings.
    Matched MeSH terms: Polymerase Chain Reaction
  9. Harvie S, Nor Aliza AR, Lela S, Razitasham S
    Trop Biomed, 2020 Jun 01;37(2):258-272.
    PMID: 33612796
    Dengue has been a public health concern for many years in Malaysia. Having knowledge on the current circulating dengue serotypes and population of vector mosquitoes is key in controlling outbreaks and future outbreak predictions. The current study reports the first study on detecting dengue virus serotypes in the Aedes mosquito population in Sibu and Miri divisions of Sarawak. Mosquito samples were collected at selected localities from September 2016 to December 2017. Localities were selected mainly focussing on urban residential areas. The mosquitoes collected comprises of the field-caught adults and immatures collected from artificial and natural water containers. Collected mosquitoes were identified to species level and screened for the presence of dengue virus using conventional reverse transcription polymerase chain reaction (RT-PCR). Dengue virus serotype 2 (DENV-2) was identified in 3 pools of field-caught female Aedes albopictus adults collected from Jalan Tong Sang, Sibu, Sibu Lake Garden, and Taman Ceria, Permyjaya, Miri, respectively. DENV-2 was also detected in one pool of adult male Ae. albopictus emerged from immatures collected from Taman Ceria, Permyjaya, Miri. The findings in this study revealed that Ae. albopictus was the main species colonizing the study areas, and the current circulating dengue virus serotype was DENV-2. This study also reports the first natural evidence of transovarial transmission of dengue in the natural population of Ae. albopictus within the study area and provides information as reference for further vector-pathogen studies.
    Matched MeSH terms: Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction
  10. He WH, Feng XX, Wu X, Zhai XH, Li YY, Zhang B, et al.
    Trop Biomed, 2020 Dec 01;37(4):871-876.
    PMID: 33612740 DOI: 10.47665/tb.37.4.871
    To evaluate the inhibitory effects of drugs on the growth of Babesia gibsoni, relative quantification real-time PCR method was developed in this study. The 18S rRNA gene was used as a target gene for the 2-ΔΔCt method analysis. Additionally, chicken RNA was added to the parasitized blood before total RNA extraction. The chicken β-actin gene was selected as an internal control gene for the 2-ΔΔCt method analysis. The 100 µL parasitized blood samples with different percentages of parasitized erythrocytes (PPEs) (3%, 1.5%, 0.75%, 0.375% and 0.1875%) were prepared for relative quantification of B. gibsoni. Regression analysis results revealed significant linear relationships between the relative quantification value and parasitemia. 18S rRNA gene expression was significantly decreased after treatment with diminazene aceturate and artesunate in vitro drug sensitivity test. This result suggested that this relative quantification real-time PCR method can be used to evaluate the effects of drug inhibition.
    Matched MeSH terms: Real-Time Polymerase Chain Reaction
  11. Chin KL, Teoh BT, Sam SS, Loong SK, Tan KK, Azizan NS, et al.
    Trop Biomed, 2022 Dec 01;39(4):518-523.
    PMID: 36602210 DOI: 10.47665/tb.39.4.005
    Zika virus (ZIKV) infection has emerged as a global health concern following epidemic outbreaks of severe neurological disorders reported in Pacific and Americas since 2016. Therefore, a rapid, sensitive and specific diagnostic test for ZIKV infection is critical for the appropriate patient management and the control of disease spread. A TaqMan minor groove binding (MGB) probe-based quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was developed based on the conserved sequence regions of 463 ZIKV NS2B genes. The designed ZIKV qRT-PCR assay was evaluated for its detection limit, strain coverage and cross-reactivity. We further assessed the clinical applicability of qRT-PCR assay for ZIKV RNA detection using a total 18 simulated clinical specimens. The detection limit of the qRT-PCR assay was 11.276 ZIKV RNA copies at the 95% probability level (probit analysis, p<= 0.05). Both Asian and African ZIKV strains were detected by the qRT-PCR assay without cross-reacting with DENV-1, DENV-2, DENV-3, DENV-4, CHIKV, JEV, LGTV, GETV and SINV. The qRT-PCR assay demonstrated a perfect agreement (k = 1.000, P < 0.001) with the reference assay; the sensitivity and specificity of the qRT-PCR assay were 100% (95% CI= 79.6-100) and 100% (95% CI= 43.9-100) respectively. The qRT-PCR assay developed in this study is a useful diagnostic tool for the broad coverage detection and quantification of both the Asian and African ZIKV strains.
    Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction; Real-Time Polymerase Chain Reaction
  12. John DV, Aryalakshmi B, Deora H, Purushottam M, Raju R, Mahadevan A, et al.
    Trop Biomed, 2022 Dec 01;39(4):489-498.
    PMID: 36602206 DOI: 10.47665/tb.39.4.002
    Despite clinical suspicion of an infection, brain abscess samples are often culture-negative in routine microbiological testing. Direct PCR of such samples enables the identification of microbes that may be fastidious, non-viable, or unculturable. Brain abscess samples (n = 217) from neurosurgical patients were subjected to broad range 16S rRNA gene PCR and sequencing for bacteria. All these samples and seven formalin-fixed paraffin-embedded tissue (FFPE) samples were subjected to species-specific 18S rRNA PCR for neurotropic free-living amoeba that harbour pathogenic bacteria. The concordance between smear and/or culture and PCR was 69%. One-third of the samples were smear- and culture-negative for bacterial agents. However, 88% of these culture-negative samples showed the presence of bacterial 16S rRNA by PCR. Sanger sequencing of 27 selected samples showed anaerobic/fastidious gram negative bacteria (GNB, 38%), facultative Streptococci (35%), and aerobic GNB (27%). Targeted metagenomics sequencing of three samples showed multiple bacterial species, including anaerobic and non-culturable bacteria. One FFPE tissue revealed the presence of Acanthamoeba 18S rRNA. None of the frozen brain abscess samples tested was positive for 18S rRNA of Acanthamoeba or Balamuthia mandrillaris. The microbial 16/18S rRNA PCR and sequencing outperformed culture in detecting anaerobes, facultative Streptococci and FLA in brain abscess samples. Genetic analyses of 16S/18S sequences, either through Sanger or metagenomic sequencing, will be an essential diagnostic technology to be included for diagnosing culture-negative brain abscess samples. Characterizing the microbiome of culture-negative brain abscess samples by molecular methods could enable detection and/or treatment of the source of infection.
    Matched MeSH terms: Polymerase Chain Reaction/methods
  13. La N, Leng M, Arunsan P, Pechdee P, Boonsuya A, Thanchonnang C, et al.
    Trop Biomed, 2023 Dec 01;40(4):383-391.
    PMID: 38308824 DOI: 10.47665/tb.40.4.002
    The southeast Asian fluke Opisthorchis viverrini remains endemic, particularly in Thailand, Lao PDR, Cambodia, Vietnam, and Myanmar. However, there is a lack of data on the prevalence of liver fluke infection in Kratie Province in northeastern Cambodia. The present study aimed to detect O. viverrini DNA in fecal specimens by using the internal transcribed spacer 2 (ITS2) region of ribosomal DNA (rDNA) based on polymerase chain reaction (PCR). The prevalence and percentage of O. viverrini infection were described by data analysis. Bivariate binary logistic regression analysis was used to look at the related prevalence of O. viverrini infection. A total of 6.89% from 377 fecal samples were found positive of O. viverrini DNA. The prevalence of O. viverrini infection was found to be higher in men (8.92%) than in women (5.45%), and to be associated more frequently with younger age groups (13.40%), illiteracy (8.74%), participation in other careers (non-specific occupations) (11.63%), and residence in the Trapaing Srae village (9.94%) of the Snuol district, Kratie Province. Age groups under 20 years old were significantly linked with O. viverrini infection, with ORadj=0.601, 95% CI=0.410-0.882, p=0.009 and significant value established at (P<0.05). This study demonstrates that O. viverrini infection is distributed in rural areas located near freshwater reservoirs. Therefore, active surveillance, clinical examination of association with hepatobiliary, cholangiocarcinoma, and health education are needed.
    Matched MeSH terms: Polymerase Chain Reaction
  14. Kutty MK, Latif B, Muslim A, Hussaini J, Daher AM, Heo CC, et al.
    Trop Anim Health Prod, 2015 Apr;47(4):751-6.
    PMID: 25740651 DOI: 10.1007/s11250-015-0789-4
    A number of methods have been used for the detection of the presence of microsarcocysts in animals, but little information exists on the value between the various methods. This study therefore examined for Sarcocystis spp. using three different methods in 105 samples of skeletal muscle collected from goats slaughtered in an abattoir in Selangor, Malaysia from January to February 2014. Three methods were used, direct light microscopy of squashed fresh muscle tissues; histological examination of fixed, sectioned, and hematoxylin and eosin (H&E)-stained samples of muscle; and molecular identification by polymerase chain reaction (PCR). Of the 105 tissue samples, 55 (52.38 %) were positive by light microscopy (LM), 46 (43.8 %) by histology, and 95 (90.48 %) by PCR. Only 29 (27.6 %) and 5 (4.76 %) samples were positive and negative, respectively, by all three methods. The cysts were elongated to a spindle shape with a mean size of 393.30 × 81.6 μm and containing banana-shaped bradyzoites of size 12.32 × 2.08 μm. The wall of the cyst was radially striated with a thickness of 2.83 μm. Samples were tested for the presence of Sarcocystis-specific 18S rRNA and were identified as Sarcocystis capracanis. Of the three methods used, the PCR test appears to be the most useful method for the diagnosis of sarcocystosis especially for species identification.
    Matched MeSH terms: Polymerase Chain Reaction/veterinary
  15. Mohd Yasin IS, Mohd Yusoff S, Mohd ZS, Abd Wahid Mohd E
    Trop Anim Health Prod, 2011 Jan;43(1):179-87.
    PMID: 20697957 DOI: 10.1007/s11250-010-9672-5
    This study was carried out to determine the antibody responses and protective capacity of an inactivated recombinant vaccine expressing the fimbrial protein of Pasteurella multocida B:2 following intranasal vaccination against hemorrhagic septicemia in goats. Goats were vaccinated intranasal with 10(6) CFU/mL of the recombinant vaccine (vaccinated group) and 10(6) CFU/mL of pET32/LIC vector without fimbrial protein (control group). All three groups were kept separated before all goats in the three groups were challenged with 10(9) CFU/mL of live pathogenic P. multocida B:2. During the course of study, both serum and lung lavage fluid were collected to evaluate the antibody levels via enzyme-linked immunosorbent assay. It was found that goats immunized with the inactivated recombinant vaccine developed a strong and significantly (p 
    Matched MeSH terms: Polymerase Chain Reaction
  16. Kumar K, Arshad SS, Toung OP, Abba Y, Selvarajah GT, Abu J, et al.
    Trop Anim Health Prod, 2019 Mar;51(3):495-506.
    PMID: 30604332 DOI: 10.1007/s11250-018-01786-x
    Flaviviruses (FVs) are arthropod-borne viruses of medical and veterinary importance. Numerous species of FVs have been isolated from various host; mainly humans, animals, ticks, and mosquitoes. Certain FVs are extremely host-specific; at the same time, some FVs can infect an extensive range of species. Based on published literatures, 11 species of FVs have been detected from diverse host species in Malaysia. In humans, dengue virus and Japanese encephalitis virus have been reported since 1901 and 1942. In animals, the Batu Cave virus, Sitiawan virus, Carey Island, Tembusu virus, Duck Tembusu virus, and Japanese encephalitis viruses were isolated from various species. In mosquitoes, Japanese encephalitis virus and Kunjin virus were isolated from Culex spp., while Zika virus and Jugra virus were isolated from Aedes spp. In ticks, the Langat virus was isolated from Ixodes spp. One of the major challenges in the diagnosis of FVs is the presence of sero-complexes as a result of cross-reactivity with one or more FV species. Subsequently, the distribution of specific FVs among humans and animals in a specific population is problematic to assess and often require comprehensive and thorough analyses. Molecular assays such as quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and digital droplet RT-PCR (ddRT-PCR) have been used for the differentiation of flavivirus infections to increase the accuracy of epidemiological data for disease surveillance, monitoring, and control. In situations where sero-complexes are common in FVs, even sensitive assays such as qRT-pCR can produce false positive results. In this write up, an overview of the various FV sero-complexes reported in Malaysia to date and the challenges faced in diagnosis of FV infections are presented.
    Matched MeSH terms: Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction
  17. Mohan Jacob D, Lee CY, Arshad SS, Selvarajah GT, Bande F, Ong BL, et al.
    Trop Anim Health Prod, 2018 Apr;50(4):733-739.
    PMID: 29243138 DOI: 10.1007/s11250-017-1489-z
    Several strains of porcine bocaviruses have been reported worldwide since their first detection in Sweden in 2009. Subsequently, the virus has been reported to be associated with gastrointestinal and respiratory signs in weaner and grower pigs. Although Malaysia is host to a self-sufficient swine livestock industry, there is no study that describes porcine bocavirus in the country. This report is the first to describe porcine bocavirus (PBoV) in Malaysian swine herds. PBoV was identified in various tissues from sick and runt pigs using the conventional PCR method with primers targeting conserved regions encoding for the nonstructural protein (NS1) gene. Out of 103 samples tested from 17 pigs, 32 samples from 15 pigs were positive for porcine bocavirus. In addition, a higher detection rate was identified from mesenteric lymph nodes (52.9%), followed by tonsil (37.0%), and lungs (33.3%). Pairwise comparison and phylogenetic analyses based on a 658-bp fragment of NS1 gene revealed that the Malaysian PBoV strains are highly similar to PBoV3 isolated in Minnesota, USA. The presence of porcine bocavirus in Malaysia and their phylogenetic bond was marked for the first time by this study. Further studies will establish the molecular epidemiology of PBoV in Malaysia and clarify pathogenicity of the local isolates.
    Matched MeSH terms: Polymerase Chain Reaction
  18. Galinski MR, Barnwell JW
    Trends Parasitol, 2009 May;25(5):200-4.
    PMID: 19345613 DOI: 10.1016/j.pt.2009.02.002
    Four human deaths caused by Plasmodium knowlesi, a simian malaria species, are stimulating a surge of public health interest and clinical vigilance in vulnerable areas of Southeast Asia. We, and other colleagues, emphasize that these cases, identified in Malaysia, are a clear warning that health facilities and clinicians must rethink the diagnosis and treatment of malaria cases presumed to be caused by a less virulent human malaria species, Plasmodium malariae.
    Matched MeSH terms: Polymerase Chain Reaction/methods
  19. CHAI SIAW YEW, CHAI SZE FAN, LESLEY MAURICE BILUNG, AHMAD SYATIR TAHAR, ROSDI KIRA
    MyJurnal
    Listeria spp. and Salmonella spp. are capable of causing food-borne outbreaks and diseases in humans. This study aimed to quantify and detect the occurrence of Listeria monocytogenes and Salmonella Typhimurium in fruit juices by utilizing Most Probable Number (MPN) in combination with Polymerase Chain Reaction (PCR). In this study, a total of 50 fruit juice samples, consisting of orange, papaya, watermelon, honeydew and apple were collected from Kota Samarahan and Kuching. Specific Polymerase Chain Reaction (PCR) assay targeting the virulence gene, hlyA gene in L. monocytogenes and fliC gene in S. Typhimurium was performed, with the expected size of 730 bp and 559 bp, respectively. MPN analysis showed that the estimated microbial loads of Listeria spp. and Salmonella spp. in all samples were more than 1100 MPN/g. However, based on the PCR analysis, none of the samples (0%) were positive for L. monocytogenes or S. Typhimurium. This study presented as a preliminary food safety screening for the occurrence of Listeria spp. and Salmonella spp. from retailed fruit juices. Hygienic practices and food safety measures should be adhered by all food vendors and restaurants in order to avoid foodborne disease outbreaks in the future.
    Matched MeSH terms: Polymerase Chain Reaction
  20. Phei-Lian, Wang, Edmund Sim, Ui Hang
    MyJurnal
    Increasing evidence of the association between ribosomal protein (RP) genes with nasopharyngeal carcinoma (NPC) have been derived from findings of their differential expression patterns in NPC cell lines. Nevertheless, expression data from a comprehensive list of RP gene family members is still lacking. This paper reports the assessment of two RP genes, eL13 and eL14, with regards to their expression patterns in several NPC cell lines (TW04, TW01, HK1, HONE1 and SUNE-1) relative to a non-malignant control (NP69). A conventional Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) assay was employed. Analysis of eL13 has never been explored before this, whereas investigation of eL14 represents an extended study. We found a general over-expression trend of eL14 in 40% (2 of 5; TW01 and HONE-1) of the NPC cell lines studied, with higher upregulated level in only one (TW01) of them. However, this pattern of expression level is not statistically significant. Expression of eL13 was not detected in any of the cell lines used. The inconsistency of these expression patterns demonstrates an elusive nature of RP activities in the malignancy of the nasopharynx.
    Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction
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