The Experts below are selected from a list of 31473 Experts worldwide ranked by ideXlab platform

Wanchai Maleewong - One of the best experts on this subject based on the ideXlab platform.

  • Rapid detection of Dirofilaria immitis in mosquito vectors and dogs using a real-time fluorescence resonance energy transfer PCR and Melting Curve Analysis.
    Veterinary parasitology, 2009
    Co-Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Viraphong Lulitanond, Wej Choochote, Piyanan Taweethavonsawat, Sudchit Chungpivat, Somboon Sangmaneedet, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR supplemented with Melting Curve Analysis for the rapid molecular detection of Dirofilaria immitis in mosquito vectors and dog blood samples was developed. This real-time FRET PCR was based on the fluorescence Melting Curve Analysis of a hybrid between an amplicon generated from the D. immitis ribosomal RNA gene sequence and specific fluorophore-labeled probes. The sensitivity, specificity, accuracy, and positive and negative predictive values of this method were all 100%. Besides being highly sensitive and specific, this PCR is fast and offers a high throughput. Therefore it is a suitable and powerful tool for the diagnosis and for epidemiological surveys of canine dirofilariasis as well as for molecular xenomonitoring of D. immitis in mosquito vectors.

  • Real-time fluorescence resonance energy transfer PCR with Melting Curve Analysis for the detection of Opisthorchis viverrini in fish intermediate hosts.
    Veterinary parasitology, 2008
    Co-Authors: Pewpan M. Intapan, Viraphong Lulitanond, Tongjit Thanchomnang, P Phongsaskulchoti, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR combined with a Melting Curve Analysis was developed for the detection of Opisthorchis viverrini in its fish intermediate host, cyprinoid fishes. Real-time FRET PCR is based on a fluorescence Melting Curve Analysis of a hybrid between an amplicon generated from a family of repeated DNA elements, the pOV-A6 specific probe sequence (Genbank Accession No. S80278), a 162 bp repeated sequence specific to O. viverrini, and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single metacercaria artificially inoculated in 30 fish samples. The O. viverrini infected fishes were distinguished from non-infected fishes and from the genomic DNA of other parasites by their Melting temperature. Sensitivity and specificity of this method were both 100% in the laboratory setting and it outperformed the microscopic method on field-collected samples as well. Melting Curve Analysis is a rapid, accurate, and sensitive alternative for the specific detection of O. viverrini infected fishes. It allows a high throughput and can be performed on small samples. The assay has not only great potential for epidemiological surveys of fish intermediate hosts but it could also be adapted as screening tool for a range of foodborne parasites in freshwater fishes.

  • Detection of Opisthorchis viverrini in infected bithynid snails by real-time fluorescence resonance energy transfer PCR-based method and Melting Curve Analysis
    Parasitology Research, 2008
    Co-Authors: Pewpan M. Intapan, Phunthira Pongsaskulchoti, Viraphong Lulitanond, Tongjit Thanchomnang, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR combined with Melting Curve Analysis was developed for the detection of Opisthorchis viverrini in experimentally infected bithynid snails, its first intermediate hosts. The test is based on the fluorescence Melting Curve Analysis of a hybrid between an amplicon from the pOV-A6-specific probe sequence, a 162-bp repeated sequence specific to O. viverrini and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single cercaria artificially introduced in a pool of 30 non-infected snails. The O. viverrini -infected snails were discriminated from non-infected snails and from genomic DNA of other parasite DNAs by their Melting temperatures. Sensitivity and specificity of this method were both 100%. Melting Curve Analysis is a sensitive alternative for the specific detection of O. viverrini -infected snails; it is rapid, allows a high throughput, and can be done on small samples. The assay not only has a high potential for epidemiological surveys of O. viverrini -infected bithynid snails, but also for the detection of cercariae infestations of natural waterways when monitoring transmission sites.

  • Rapid detection of Brugia malayi in mosquito vectors using a real-time fluorescence resonance energy transfer PCR and Melting Curve Analysis.
    The American journal of tropical medicine and hygiene, 2008
    Co-Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Viraphong Lulitanond, Wej Choochote, Anonglak Manjai, Thidarat K. Prasongdee, Wanchai Maleewong
    Abstract:

    We developed real-time fluorescence resonance energy transfer (FRET) polymerase chain reaction (PCR) combined with Melting Curve Analysis for detection of Brugia malayi DNA in blood-fed mosquitoes. Real-time FRET PCR is based on a fluorescence Melting Curve Analysis of hybrid formed between amplicons generated from a family of repeated DNA element, 153-bp HhaI repeated sequence, specific to genus Brugia and specific fluorophore-labeled probes. The B. malayi-infected mosquitoes were differentiated from Wuchereria bancrofti-infected and uninfected mosquitoes and from genomic DNA of Dirofilaria immitis- and Plasmodium falciparum-infected human red blood cells and human leukocytes by their Melting temperature. Sensitivity and specificity were both 100%. Melting Curve Analysis produces a rapid, accurate, and sensitive alternative for specific detection of B. malayi in mosquitoes, allows high throughput, and can be performed on small samples. This method has the potential for endemic area mapping or monitoring effect of brugian filariasis mass treatment programs.

  • Rapid detection of Wuchereria bancrofti in mosquitoes by LightCycler polymerase chain reaction and Melting Curve Analysis.
    Parasitology research, 2004
    Co-Authors: Viraphong Lulitanond, Pewpan M. Intapan, V. Pipitgool, Wej Choochote, Wanchai Maleewong
    Abstract:

    A LightCycler real-time polymerase chain reaction (PCR) assay was developed to detect Wuchereria bancrofti DNA in blood-fed mosquitoes. The assay is based on fluorescence Melting Curve Analysis of the PCR product generated from a family of repeated DNA elements: the 182 bp SspI repeat, specific to the genus Wuchereria. According to the Melting temperature, W. bancrofti infected-mosquitoes were differentiated from Brugia malayi-infected and non-infected mosquitoes as well as from genomic DNA of Dirofilaria immitis and human DNA. The method proved to be 100% sensitive in all W. bancrofti-infected mosquitoes. Melting Curve Analysis offers a rapid alternative for the specific detection of W. bancrofti in mosquitoes. It is very accurate and sensitive, allows a high throughput and can be performed on very small samples. The method therefore has great potential for application in epidemiological studies.

Pewpan M. Intapan - One of the best experts on this subject based on the ideXlab platform.

  • Detection of Ehrlichia canis in canine blood samples by real-time fluorescence resonance energy transfer (FRET) PCR and Melting Curve Analysis.
    The Southeast Asian journal of tropical medicine and public health, 2014
    Co-Authors: Amornmas Kongklieng, Pewpan M. Intapan, Viraphong Lulitanond, Tongjit Thanchomnang, Thidarut Boonmars, Penchom Janwan, Oranuch Sanpool, Piyanan Taweethavonsawat, Sudchit Chungpivat, Nimit Morakote
    Abstract:

    Ehrlichia canis is a small pleomorphic gram-negative, coccoid, obligatory intracellular bacterium and the cause of canine monocytic ehrlichiosis. A real-time fluorescence resonance energy transfer polymerase chain reaction (real-time FRET PCR) coupled with Melting Curve Analysis was established for detection of E. canis infection in canine blood samples. The VirB9 gene was amplified using one pair of primers and the Melting Curve Analysis was generated by heating the hybridizing probes and amplified products. Eight E. canis-infected dog blood samples were initially identified using the Giemsa staining/microscopic method followed by conventional PCR (cPCR)/Sanger sequencing for confirmation. The sensitivity and specificity of the real-time FRET PCR detection were 87.5% and 100%, respectively and the limit of detection was 6.6 x 10(3) copies of positive E. canis control plasmids. The real-time FRET PCR with Melting Curve Analysis reported here is better than microscopic visualization or cPCR because the method is not affected by the false bias inherent in the microscopic method. Furthermore, many samples can be processed rapidly at the same time. This convenient tool is beneficial as an alternative assay for the epidemiologic study of canine ehrlichiosis as well as for eradication of these organisms in prevention and control programs in endemic areas.

  • Rapid detection of Dirofilaria immitis in mosquito vectors and dogs using a real-time fluorescence resonance energy transfer PCR and Melting Curve Analysis.
    Veterinary parasitology, 2009
    Co-Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Viraphong Lulitanond, Wej Choochote, Piyanan Taweethavonsawat, Sudchit Chungpivat, Somboon Sangmaneedet, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR supplemented with Melting Curve Analysis for the rapid molecular detection of Dirofilaria immitis in mosquito vectors and dog blood samples was developed. This real-time FRET PCR was based on the fluorescence Melting Curve Analysis of a hybrid between an amplicon generated from the D. immitis ribosomal RNA gene sequence and specific fluorophore-labeled probes. The sensitivity, specificity, accuracy, and positive and negative predictive values of this method were all 100%. Besides being highly sensitive and specific, this PCR is fast and offers a high throughput. Therefore it is a suitable and powerful tool for the diagnosis and for epidemiological surveys of canine dirofilariasis as well as for molecular xenomonitoring of D. immitis in mosquito vectors.

  • Real-time fluorescence resonance energy transfer PCR with Melting Curve Analysis for the detection of Opisthorchis viverrini in fish intermediate hosts.
    Veterinary parasitology, 2008
    Co-Authors: Pewpan M. Intapan, Viraphong Lulitanond, Tongjit Thanchomnang, P Phongsaskulchoti, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR combined with a Melting Curve Analysis was developed for the detection of Opisthorchis viverrini in its fish intermediate host, cyprinoid fishes. Real-time FRET PCR is based on a fluorescence Melting Curve Analysis of a hybrid between an amplicon generated from a family of repeated DNA elements, the pOV-A6 specific probe sequence (Genbank Accession No. S80278), a 162 bp repeated sequence specific to O. viverrini, and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single metacercaria artificially inoculated in 30 fish samples. The O. viverrini infected fishes were distinguished from non-infected fishes and from the genomic DNA of other parasites by their Melting temperature. Sensitivity and specificity of this method were both 100% in the laboratory setting and it outperformed the microscopic method on field-collected samples as well. Melting Curve Analysis is a rapid, accurate, and sensitive alternative for the specific detection of O. viverrini infected fishes. It allows a high throughput and can be performed on small samples. The assay has not only great potential for epidemiological surveys of fish intermediate hosts but it could also be adapted as screening tool for a range of foodborne parasites in freshwater fishes.

  • Detection of Opisthorchis viverrini in infected bithynid snails by real-time fluorescence resonance energy transfer PCR-based method and Melting Curve Analysis
    Parasitology Research, 2008
    Co-Authors: Pewpan M. Intapan, Phunthira Pongsaskulchoti, Viraphong Lulitanond, Tongjit Thanchomnang, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR combined with Melting Curve Analysis was developed for the detection of Opisthorchis viverrini in experimentally infected bithynid snails, its first intermediate hosts. The test is based on the fluorescence Melting Curve Analysis of a hybrid between an amplicon from the pOV-A6-specific probe sequence, a 162-bp repeated sequence specific to O. viverrini and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single cercaria artificially introduced in a pool of 30 non-infected snails. The O. viverrini -infected snails were discriminated from non-infected snails and from genomic DNA of other parasite DNAs by their Melting temperatures. Sensitivity and specificity of this method were both 100%. Melting Curve Analysis is a sensitive alternative for the specific detection of O. viverrini -infected snails; it is rapid, allows a high throughput, and can be done on small samples. The assay not only has a high potential for epidemiological surveys of O. viverrini -infected bithynid snails, but also for the detection of cercariae infestations of natural waterways when monitoring transmission sites.

  • Rapid detection of Brugia malayi in mosquito vectors using a real-time fluorescence resonance energy transfer PCR and Melting Curve Analysis.
    The American journal of tropical medicine and hygiene, 2008
    Co-Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Viraphong Lulitanond, Wej Choochote, Anonglak Manjai, Thidarat K. Prasongdee, Wanchai Maleewong
    Abstract:

    We developed real-time fluorescence resonance energy transfer (FRET) polymerase chain reaction (PCR) combined with Melting Curve Analysis for detection of Brugia malayi DNA in blood-fed mosquitoes. Real-time FRET PCR is based on a fluorescence Melting Curve Analysis of hybrid formed between amplicons generated from a family of repeated DNA element, 153-bp HhaI repeated sequence, specific to genus Brugia and specific fluorophore-labeled probes. The B. malayi-infected mosquitoes were differentiated from Wuchereria bancrofti-infected and uninfected mosquitoes and from genomic DNA of Dirofilaria immitis- and Plasmodium falciparum-infected human red blood cells and human leukocytes by their Melting temperature. Sensitivity and specificity were both 100%. Melting Curve Analysis produces a rapid, accurate, and sensitive alternative for specific detection of B. malayi in mosquitoes, allows high throughput, and can be performed on small samples. This method has the potential for endemic area mapping or monitoring effect of brugian filariasis mass treatment programs.

Viraphong Lulitanond - One of the best experts on this subject based on the ideXlab platform.

  • Detection of Ehrlichia canis in canine blood samples by real-time fluorescence resonance energy transfer (FRET) PCR and Melting Curve Analysis.
    The Southeast Asian journal of tropical medicine and public health, 2014
    Co-Authors: Amornmas Kongklieng, Pewpan M. Intapan, Viraphong Lulitanond, Tongjit Thanchomnang, Thidarut Boonmars, Penchom Janwan, Oranuch Sanpool, Piyanan Taweethavonsawat, Sudchit Chungpivat, Nimit Morakote
    Abstract:

    Ehrlichia canis is a small pleomorphic gram-negative, coccoid, obligatory intracellular bacterium and the cause of canine monocytic ehrlichiosis. A real-time fluorescence resonance energy transfer polymerase chain reaction (real-time FRET PCR) coupled with Melting Curve Analysis was established for detection of E. canis infection in canine blood samples. The VirB9 gene was amplified using one pair of primers and the Melting Curve Analysis was generated by heating the hybridizing probes and amplified products. Eight E. canis-infected dog blood samples were initially identified using the Giemsa staining/microscopic method followed by conventional PCR (cPCR)/Sanger sequencing for confirmation. The sensitivity and specificity of the real-time FRET PCR detection were 87.5% and 100%, respectively and the limit of detection was 6.6 x 10(3) copies of positive E. canis control plasmids. The real-time FRET PCR with Melting Curve Analysis reported here is better than microscopic visualization or cPCR because the method is not affected by the false bias inherent in the microscopic method. Furthermore, many samples can be processed rapidly at the same time. This convenient tool is beneficial as an alternative assay for the epidemiologic study of canine ehrlichiosis as well as for eradication of these organisms in prevention and control programs in endemic areas.

  • Rapid detection of Dirofilaria immitis in mosquito vectors and dogs using a real-time fluorescence resonance energy transfer PCR and Melting Curve Analysis.
    Veterinary parasitology, 2009
    Co-Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Viraphong Lulitanond, Wej Choochote, Piyanan Taweethavonsawat, Sudchit Chungpivat, Somboon Sangmaneedet, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR supplemented with Melting Curve Analysis for the rapid molecular detection of Dirofilaria immitis in mosquito vectors and dog blood samples was developed. This real-time FRET PCR was based on the fluorescence Melting Curve Analysis of a hybrid between an amplicon generated from the D. immitis ribosomal RNA gene sequence and specific fluorophore-labeled probes. The sensitivity, specificity, accuracy, and positive and negative predictive values of this method were all 100%. Besides being highly sensitive and specific, this PCR is fast and offers a high throughput. Therefore it is a suitable and powerful tool for the diagnosis and for epidemiological surveys of canine dirofilariasis as well as for molecular xenomonitoring of D. immitis in mosquito vectors.

  • Real-time fluorescence resonance energy transfer PCR with Melting Curve Analysis for the detection of Opisthorchis viverrini in fish intermediate hosts.
    Veterinary parasitology, 2008
    Co-Authors: Pewpan M. Intapan, Viraphong Lulitanond, Tongjit Thanchomnang, P Phongsaskulchoti, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR combined with a Melting Curve Analysis was developed for the detection of Opisthorchis viverrini in its fish intermediate host, cyprinoid fishes. Real-time FRET PCR is based on a fluorescence Melting Curve Analysis of a hybrid between an amplicon generated from a family of repeated DNA elements, the pOV-A6 specific probe sequence (Genbank Accession No. S80278), a 162 bp repeated sequence specific to O. viverrini, and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single metacercaria artificially inoculated in 30 fish samples. The O. viverrini infected fishes were distinguished from non-infected fishes and from the genomic DNA of other parasites by their Melting temperature. Sensitivity and specificity of this method were both 100% in the laboratory setting and it outperformed the microscopic method on field-collected samples as well. Melting Curve Analysis is a rapid, accurate, and sensitive alternative for the specific detection of O. viverrini infected fishes. It allows a high throughput and can be performed on small samples. The assay has not only great potential for epidemiological surveys of fish intermediate hosts but it could also be adapted as screening tool for a range of foodborne parasites in freshwater fishes.

  • Detection of Opisthorchis viverrini in infected bithynid snails by real-time fluorescence resonance energy transfer PCR-based method and Melting Curve Analysis
    Parasitology Research, 2008
    Co-Authors: Pewpan M. Intapan, Phunthira Pongsaskulchoti, Viraphong Lulitanond, Tongjit Thanchomnang, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR combined with Melting Curve Analysis was developed for the detection of Opisthorchis viverrini in experimentally infected bithynid snails, its first intermediate hosts. The test is based on the fluorescence Melting Curve Analysis of a hybrid between an amplicon from the pOV-A6-specific probe sequence, a 162-bp repeated sequence specific to O. viverrini and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single cercaria artificially introduced in a pool of 30 non-infected snails. The O. viverrini -infected snails were discriminated from non-infected snails and from genomic DNA of other parasite DNAs by their Melting temperatures. Sensitivity and specificity of this method were both 100%. Melting Curve Analysis is a sensitive alternative for the specific detection of O. viverrini -infected snails; it is rapid, allows a high throughput, and can be done on small samples. The assay not only has a high potential for epidemiological surveys of O. viverrini -infected bithynid snails, but also for the detection of cercariae infestations of natural waterways when monitoring transmission sites.

  • Rapid detection of Brugia malayi in mosquito vectors using a real-time fluorescence resonance energy transfer PCR and Melting Curve Analysis.
    The American journal of tropical medicine and hygiene, 2008
    Co-Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Viraphong Lulitanond, Wej Choochote, Anonglak Manjai, Thidarat K. Prasongdee, Wanchai Maleewong
    Abstract:

    We developed real-time fluorescence resonance energy transfer (FRET) polymerase chain reaction (PCR) combined with Melting Curve Analysis for detection of Brugia malayi DNA in blood-fed mosquitoes. Real-time FRET PCR is based on a fluorescence Melting Curve Analysis of hybrid formed between amplicons generated from a family of repeated DNA element, 153-bp HhaI repeated sequence, specific to genus Brugia and specific fluorophore-labeled probes. The B. malayi-infected mosquitoes were differentiated from Wuchereria bancrofti-infected and uninfected mosquitoes and from genomic DNA of Dirofilaria immitis- and Plasmodium falciparum-infected human red blood cells and human leukocytes by their Melting temperature. Sensitivity and specificity were both 100%. Melting Curve Analysis produces a rapid, accurate, and sensitive alternative for specific detection of B. malayi in mosquitoes, allows high throughput, and can be performed on small samples. This method has the potential for endemic area mapping or monitoring effect of brugian filariasis mass treatment programs.

Tongjit Thanchomnang - One of the best experts on this subject based on the ideXlab platform.

  • Detection of Ehrlichia canis in canine blood samples by real-time fluorescence resonance energy transfer (FRET) PCR and Melting Curve Analysis.
    The Southeast Asian journal of tropical medicine and public health, 2014
    Co-Authors: Amornmas Kongklieng, Pewpan M. Intapan, Viraphong Lulitanond, Tongjit Thanchomnang, Thidarut Boonmars, Penchom Janwan, Oranuch Sanpool, Piyanan Taweethavonsawat, Sudchit Chungpivat, Nimit Morakote
    Abstract:

    Ehrlichia canis is a small pleomorphic gram-negative, coccoid, obligatory intracellular bacterium and the cause of canine monocytic ehrlichiosis. A real-time fluorescence resonance energy transfer polymerase chain reaction (real-time FRET PCR) coupled with Melting Curve Analysis was established for detection of E. canis infection in canine blood samples. The VirB9 gene was amplified using one pair of primers and the Melting Curve Analysis was generated by heating the hybridizing probes and amplified products. Eight E. canis-infected dog blood samples were initially identified using the Giemsa staining/microscopic method followed by conventional PCR (cPCR)/Sanger sequencing for confirmation. The sensitivity and specificity of the real-time FRET PCR detection were 87.5% and 100%, respectively and the limit of detection was 6.6 x 10(3) copies of positive E. canis control plasmids. The real-time FRET PCR with Melting Curve Analysis reported here is better than microscopic visualization or cPCR because the method is not affected by the false bias inherent in the microscopic method. Furthermore, many samples can be processed rapidly at the same time. This convenient tool is beneficial as an alternative assay for the epidemiologic study of canine ehrlichiosis as well as for eradication of these organisms in prevention and control programs in endemic areas.

  • Rapid detection of Dirofilaria immitis in mosquito vectors and dogs using a real-time fluorescence resonance energy transfer PCR and Melting Curve Analysis.
    Veterinary parasitology, 2009
    Co-Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Viraphong Lulitanond, Wej Choochote, Piyanan Taweethavonsawat, Sudchit Chungpivat, Somboon Sangmaneedet, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR supplemented with Melting Curve Analysis for the rapid molecular detection of Dirofilaria immitis in mosquito vectors and dog blood samples was developed. This real-time FRET PCR was based on the fluorescence Melting Curve Analysis of a hybrid between an amplicon generated from the D. immitis ribosomal RNA gene sequence and specific fluorophore-labeled probes. The sensitivity, specificity, accuracy, and positive and negative predictive values of this method were all 100%. Besides being highly sensitive and specific, this PCR is fast and offers a high throughput. Therefore it is a suitable and powerful tool for the diagnosis and for epidemiological surveys of canine dirofilariasis as well as for molecular xenomonitoring of D. immitis in mosquito vectors.

  • Real-time fluorescence resonance energy transfer PCR with Melting Curve Analysis for the detection of Opisthorchis viverrini in fish intermediate hosts.
    Veterinary parasitology, 2008
    Co-Authors: Pewpan M. Intapan, Viraphong Lulitanond, Tongjit Thanchomnang, P Phongsaskulchoti, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR combined with a Melting Curve Analysis was developed for the detection of Opisthorchis viverrini in its fish intermediate host, cyprinoid fishes. Real-time FRET PCR is based on a fluorescence Melting Curve Analysis of a hybrid between an amplicon generated from a family of repeated DNA elements, the pOV-A6 specific probe sequence (Genbank Accession No. S80278), a 162 bp repeated sequence specific to O. viverrini, and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single metacercaria artificially inoculated in 30 fish samples. The O. viverrini infected fishes were distinguished from non-infected fishes and from the genomic DNA of other parasites by their Melting temperature. Sensitivity and specificity of this method were both 100% in the laboratory setting and it outperformed the microscopic method on field-collected samples as well. Melting Curve Analysis is a rapid, accurate, and sensitive alternative for the specific detection of O. viverrini infected fishes. It allows a high throughput and can be performed on small samples. The assay has not only great potential for epidemiological surveys of fish intermediate hosts but it could also be adapted as screening tool for a range of foodborne parasites in freshwater fishes.

  • Detection of Opisthorchis viverrini in infected bithynid snails by real-time fluorescence resonance energy transfer PCR-based method and Melting Curve Analysis
    Parasitology Research, 2008
    Co-Authors: Pewpan M. Intapan, Phunthira Pongsaskulchoti, Viraphong Lulitanond, Tongjit Thanchomnang, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR combined with Melting Curve Analysis was developed for the detection of Opisthorchis viverrini in experimentally infected bithynid snails, its first intermediate hosts. The test is based on the fluorescence Melting Curve Analysis of a hybrid between an amplicon from the pOV-A6-specific probe sequence, a 162-bp repeated sequence specific to O. viverrini and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single cercaria artificially introduced in a pool of 30 non-infected snails. The O. viverrini -infected snails were discriminated from non-infected snails and from genomic DNA of other parasite DNAs by their Melting temperatures. Sensitivity and specificity of this method were both 100%. Melting Curve Analysis is a sensitive alternative for the specific detection of O. viverrini -infected snails; it is rapid, allows a high throughput, and can be done on small samples. The assay not only has a high potential for epidemiological surveys of O. viverrini -infected bithynid snails, but also for the detection of cercariae infestations of natural waterways when monitoring transmission sites.

  • Rapid detection of Brugia malayi in mosquito vectors using a real-time fluorescence resonance energy transfer PCR and Melting Curve Analysis.
    The American journal of tropical medicine and hygiene, 2008
    Co-Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Viraphong Lulitanond, Wej Choochote, Anonglak Manjai, Thidarat K. Prasongdee, Wanchai Maleewong
    Abstract:

    We developed real-time fluorescence resonance energy transfer (FRET) polymerase chain reaction (PCR) combined with Melting Curve Analysis for detection of Brugia malayi DNA in blood-fed mosquitoes. Real-time FRET PCR is based on a fluorescence Melting Curve Analysis of hybrid formed between amplicons generated from a family of repeated DNA element, 153-bp HhaI repeated sequence, specific to genus Brugia and specific fluorophore-labeled probes. The B. malayi-infected mosquitoes were differentiated from Wuchereria bancrofti-infected and uninfected mosquitoes and from genomic DNA of Dirofilaria immitis- and Plasmodium falciparum-infected human red blood cells and human leukocytes by their Melting temperature. Sensitivity and specificity were both 100%. Melting Curve Analysis produces a rapid, accurate, and sensitive alternative for specific detection of B. malayi in mosquitoes, allows high throughput, and can be performed on small samples. This method has the potential for endemic area mapping or monitoring effect of brugian filariasis mass treatment programs.

Wej Choochote - One of the best experts on this subject based on the ideXlab platform.

  • Rapid detection of Dirofilaria immitis in mosquito vectors and dogs using a real-time fluorescence resonance energy transfer PCR and Melting Curve Analysis.
    Veterinary parasitology, 2009
    Co-Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Viraphong Lulitanond, Wej Choochote, Piyanan Taweethavonsawat, Sudchit Chungpivat, Somboon Sangmaneedet, Wanchai Maleewong
    Abstract:

    A real-time fluorescence resonance energy transfer (FRET) PCR supplemented with Melting Curve Analysis for the rapid molecular detection of Dirofilaria immitis in mosquito vectors and dog blood samples was developed. This real-time FRET PCR was based on the fluorescence Melting Curve Analysis of a hybrid between an amplicon generated from the D. immitis ribosomal RNA gene sequence and specific fluorophore-labeled probes. The sensitivity, specificity, accuracy, and positive and negative predictive values of this method were all 100%. Besides being highly sensitive and specific, this PCR is fast and offers a high throughput. Therefore it is a suitable and powerful tool for the diagnosis and for epidemiological surveys of canine dirofilariasis as well as for molecular xenomonitoring of D. immitis in mosquito vectors.

  • Rapid detection of Brugia malayi in mosquito vectors using a real-time fluorescence resonance energy transfer PCR and Melting Curve Analysis.
    The American journal of tropical medicine and hygiene, 2008
    Co-Authors: Tongjit Thanchomnang, Pewpan M. Intapan, Viraphong Lulitanond, Wej Choochote, Anonglak Manjai, Thidarat K. Prasongdee, Wanchai Maleewong
    Abstract:

    We developed real-time fluorescence resonance energy transfer (FRET) polymerase chain reaction (PCR) combined with Melting Curve Analysis for detection of Brugia malayi DNA in blood-fed mosquitoes. Real-time FRET PCR is based on a fluorescence Melting Curve Analysis of hybrid formed between amplicons generated from a family of repeated DNA element, 153-bp HhaI repeated sequence, specific to genus Brugia and specific fluorophore-labeled probes. The B. malayi-infected mosquitoes were differentiated from Wuchereria bancrofti-infected and uninfected mosquitoes and from genomic DNA of Dirofilaria immitis- and Plasmodium falciparum-infected human red blood cells and human leukocytes by their Melting temperature. Sensitivity and specificity were both 100%. Melting Curve Analysis produces a rapid, accurate, and sensitive alternative for specific detection of B. malayi in mosquitoes, allows high throughput, and can be performed on small samples. This method has the potential for endemic area mapping or monitoring effect of brugian filariasis mass treatment programs.

  • Rapid detection of Wuchereria bancrofti in mosquitoes by LightCycler polymerase chain reaction and Melting Curve Analysis.
    Parasitology research, 2004
    Co-Authors: Viraphong Lulitanond, Pewpan M. Intapan, V. Pipitgool, Wej Choochote, Wanchai Maleewong
    Abstract:

    A LightCycler real-time polymerase chain reaction (PCR) assay was developed to detect Wuchereria bancrofti DNA in blood-fed mosquitoes. The assay is based on fluorescence Melting Curve Analysis of the PCR product generated from a family of repeated DNA elements: the 182 bp SspI repeat, specific to the genus Wuchereria. According to the Melting temperature, W. bancrofti infected-mosquitoes were differentiated from Brugia malayi-infected and non-infected mosquitoes as well as from genomic DNA of Dirofilaria immitis and human DNA. The method proved to be 100% sensitive in all W. bancrofti-infected mosquitoes. Melting Curve Analysis offers a rapid alternative for the specific detection of W. bancrofti in mosquitoes. It is very accurate and sensitive, allows a high throughput and can be performed on very small samples. The method therefore has great potential for application in epidemiological studies.