A dual colour fluorescence in situ hybridization (FISH) assay for identifying the zoonotic malaria parasite Plasmodium knowlesi with a potential application for the specific diagnosis of knowlesi malaria in peripheral-level laboratories of Southeast Asia

Abstract BackgroundThe vectors for Plasmodium knowlesi, a significant cause of human malaria in Southeast Asia, identified previously in nature all belong to the Anopheles Leucosphyrus Group. Only one study has been previously undertaken in Sarawak, Malaysian Borneo, to identify vectors of P. knowlesi, where Anopheles latens was incriminated as the vector in Kapit, central Sarawak. A study was therefore undertaken to identify malaria vectors in a different location in Sarawak. MethodsMosquitoes found landing on humans and resting on leaves over a 5-day period at two sites in the Lawas District of northern Sarawak were collected and identified. DNA samples extracted from salivary glands of anophelines were subjected to nested PCR malaria-detection assays. The small sub-unit ribosomal RNA (SSUrRNA) genes of Plasmodium, and the internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome c oxidase subunit 1 (CO1) sequences of the mosquitoes were derived from the Plasmodium-positive samples for phylogenetic analyses. ResultsA total of 65 anophelines and 127 culicines were collected. By PCR, six An. balabacensis and five An. barbirostris were found to have single P. knowlesi infecions while three other An. balabacensis had either single, double or triple infections with P. inui, P. fieldi, P. cynomolgi and P. knowlesi. Phylogenetic analyses of the Plasmodium SSUrRNA genes confirmed 3 An. barbirostris and 3 An. balabacensis with single P. knowlesi infections, while 3 other An. balabacensis had two or more Plasmodium species of P. inui, P. knowlesi, P. cynomolgi and possibly novel species of Plasmodium. Phylogenies inferred from the ITS2 and CO1 sequences of An. balabacensis and An. barbirostris indicate that the former is genetically indistinguishable from An. balabacensis in Borneo while the latter is a novel sibling species belonging to the Anopheles Barbirostris Subgroup. ConclusionsNew vectors for P. knowlesi in Sarawak were identified, including An. barbirostris, which is a species that does not belong to the Anopheles Leucosphyrus Group.

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Comparison of dual-colour chromogenic in-situ hybridization and fluorescence in-situ hybridization in the assessment of HER2 gene amplification in breast carcinoma

Histopathology ◽

10.1111/j.1365-2559.2010.03737.x ◽

2011 ◽

Vol 58 (2) ◽

pp. 319-321 ◽

Cited By ~ 2

Author(s):

Susan Conlon ◽

Aoife Colgan ◽

David Allen ◽

Anthony O’Grady ◽

Elaine W Kay

Keyword(s):

In Situ Hybridization ◽

Breast Carcinoma ◽

Fluorescence In Situ Hybridization ◽

Gene Amplification ◽

Her2 Gene ◽

Her2 Gene Amplification ◽

Chromogenic In Situ Hybridization ◽

Dual Colour

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Environmental risk factors and exposure to the zoonotic malaria parasite Plasmodium knowlesi across northern Sabah, Malaysia: a population-based cross-sectional survey

The Lancet Planetary Health ◽

10.1016/s2542-5196(19)30045-2 ◽

2019 ◽

Vol 3 (4) ◽

pp. e179-e186 ◽

Cited By ~ 11

Author(s):

Kimberly M Fornace ◽

Paddy M Brock ◽

Tommy R Abidin ◽

Lynn Grignard ◽

Lou S Herman ◽

...

Keyword(s):

Risk Factors ◽

Environmental Risk ◽

Malaria Parasite ◽

Plasmodium Knowlesi ◽

Population Based ◽

Environmental Risk Factors ◽

Cross Sectional Survey ◽

Cross Sectional ◽

Zoonotic Malaria ◽

Parasite Plasmodium

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Validation of sperm sexing in the cattle (Bos taurus) by dual colour fluorescence in situ hybridization

Journal of Animal Breeding and Genetics ◽

10.1111/j.1439-0388.2005.00488.x ◽

2005 ◽

Vol 122 (s1) ◽

pp. 22-27 ◽

Cited By ~ 14

Author(s):

F.A. Habermann ◽

A. Winter ◽

I. Olsaker ◽

P. Reichert ◽

R. Fries

Keyword(s):

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Bos Taurus ◽

Dual Colour

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Disomy frequency estimated by multicolour fluorescence in situ hybridization, degree of nuclear maturity and teratozoospermia in human spermatozoa

Reproduction ◽

10.1530/rep.0.1210783 ◽

2001 ◽

pp. 783-789 ◽

Cited By ~ 16

Author(s):

F Morel ◽

C Roux ◽

JL Bresson

Keyword(s):

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Semen Analysis ◽

Human Spermatozoa ◽

Aniline Blue ◽

Blue Staining ◽

The Difference ◽

Dual Colour ◽

Aniline Blue Staining

The aim of this study was to determine whether sperm morphology and nuclear maturity are associated with an increase in the prevalence of disomy in human spermatozoa. Semen samples were obtained from 60 patients selected at random from a population of men undergoing semen analysis as part of consultation for infertility. Semen analysis and aniline blue staining were carried out on each ejaculate to assess nuclear maturity. Disomy frequencies were determined using 15-18 dual colour and X-Y-8 multicolour fluorescence in situ hybridization on the four groups of samples with the five lowest and the five highest teratozoospermia values (groups 1 and 2), and the five lowest and the five highest percentages of aniline blue staining (groups 3 and 4). The prevalence of autosomal disomy was significantly higher in group 4 compared with group 3, but differences between groups 1 and 2 were not significant. No significant differences in the prevalence of gonosomal disomies or diploidies were observed among the groups. These results indicate a link between chromosomal meiotic segregation and the dynamic process of nucleoproteins during gametogenesis. The difference observed between the frequency of autosomal and gonosomal disomy using aniline blue staining was unexpected and may be due to the extended isolation of the gonosomes in the heterochromatic body.

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