- INTRODUCTION
- MATERIALS AND METHODS
- Detection of microfilaremia and filarial antigenemia.
- DNA isolation.
- Mosquito collection and DNA extraction.
- Real-time PCR assays for detection of W. bancrofti and Wolbachia DNA.
- C-PCR assay to detect Wuchereria bancrofti DNA in mosquitoes.
- Detection of W. bancrofti microfilariae in night blood samples.
- Ethical clearance.
- Data analysis.
- RESULTS
- Detection of W. bancrofti and Wolbachia DNA using real-time PCR.
- Specificity of W. bancrofti DNA detection by real-time PCR.
- Sensitivity of real-time PCR for detecting W. bancrofti DNA in dried blood from filariasis antigen card tests.
- Sensitivity of real-time PCR and C-PCR for detecting W. bancrofti DNA in Cx. pipiens and An. punctulatus mosquitoes.
- Effects of MDA on mosquito infection rates.
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Grove DI, Warren KS, Mahmoud AA, 1975. Algorithms in the diagnosis and management of exotic diseases. VI. The filariases. J Infect Dis 132 :340–352.
-
2
Weil GJ, Lammie PJ, Weiss N, 1997. The ICT filariasis test: a rapid-format antigen test for diagnosis of bancroftian filariasis. Parasitol Today 13 :401–404.
-
3
Lammie P, Weil G, Rahmah N, Kaliraj P, Steel C, Goodman D, Lakshmikanthan V, Ottesen E, 2004. Recombinant antigen based assays for the diagnosis and surveillance of lymphatic filariasis - a multicenter trial. Filaria J 3 :9.
-
4
Williams S, Laney S, Bierwert L, Saunders L, Boakye D, Fischer P, Goodman D, Helmy H, Hoti S, Vasuki V, Lammie P, Plichart C, Ramzy R, Ottesen E, 2002. Development and standardization of a rapid PCR-based method for the detection of Wuchereria bancrofti in mosquitoes for xenomonitoring the human prevalence of bancroftian filariasis. Ann Trop Med Parasitol 96 :S41–S46.
-
5
Ramzy R, Farid H, Kamal H, Ibrahim G, Morsy Z, Faris R, Weil G, 1997. A polymerase chain reaction-based assay for detection of Wuchereria bancrofti in human blood and Culex pipiens. Trans Roy Soc Trop Med Hyg 91 :156–160.
-
6
Zhong M, McCarthy J, Bierwert L, Lizotte-Waniewski M, Chanteau S, Nutman T, Ottesen E, Williams S, 1996. A polymerase chain reaction assay for detection of the parasite Wuchereria bancrofti in human blood samples. Am J Trop Med Hyg 54 :357–363.
-
7
McCarthy J, Zhong M, Gopinath R, Ottesen E, Williams S, Nutman T, 1996. Evaluation of a polymerase chain reaction-based assay for diagnosis of Wuchereria bancrofti infection. J Infect Dis 173 :1510–1514.
-
8
Fischer P, Liu X, Lizotte-Waniewski M, Kamal I, Ramzy R, Williams S, 1999. Development of a quantitative, competitive polymerase chain reaction-enzyme-linked immunosorbent assay for the detection of Wuchereria bancrofti DNA. Parasit Res 85 :176–183.
-
9
Abbasi I, Hamburger J, Githure J, Ochola J, Agure R, Koech D, Ramzy R, Gad A, Williams S, 1996. Detection of Wuchereria bancrofti DNA in patients sputum by the polymerase chain reaction. Trans Roy Soc Trop Med Hyg 90 :531–532.
-
10
Kluber S, Supali T, Williams SA, Liebau E, Fischer P, 2001. Rapid PCR-based detection of Brugia malayi DNA from blood spots by DNA detection test strips. Trans R Soc Trop Med Hyg 95 :169–170.
-
11
Bockarie MJ, Fischer P, Williams SA, Zimmerman PA, Griffin L, Alpers MP, Kazura JW, 2000. Application of a polymerase chain reaction-ELISA to detect Wuchereria bancrofti in pools of wild-caught Anopheles punctulatus in a filariasis control area in Papua New Guinea. Am J Trop Med Hyg 62 :363–367.
-
12
Chanteau S, Luquiaud P, Failloux AB, Williams SA, 1994. Detection of Wuchereria bancrofti larvae in pools of mosquitoes by the polymerase chain reaction. Trans R Soc Trop Med Hyg 88 :665–666.
-
13
Goodman DS, Orelus JN, Roberts JM, Lammie PJ, Streit TG, 2003. PCR and Mosquito dissection as tools to monitor filarial infection levels following mass treatment. Filaria J 2 :11.
-
14
Farid HA, Hammad RE, Hassan MM, Morsy ZS, Kamal IH, Weil GJ, Ramzy RM, 2001. Detection of Wuchereria bancrofti in mosquitoes by the polymerase chain reaction: a potentially useful tool for large-scale control programmes. Trans R Soc Trop Med Hyg 95 :29–32.
-
15
Vasuki V, Hoti SL, Sadanandane C, Jambulingam P, 2003. A simple and rapid DNA extraction method for the detection of Wuchereria bancrofti infection in the vector mosquito, Culex quinquefasciatus by Ssp I PCR assay. Acta Trop 86 :109–114.
-
16
Molyneux D, 2003. Lymphatic filariasis (elephantiasis) elimination: a public health success and development opportunity. Filaria J 2 :13.
-
17
Ottesen EA, 2002. Major progress toward eliminating lymphatic filariasis. N Engl J Med 347 :1885–1886.
-
18
World Health Organization, 2005. Global programme to eliminate lymphatic filariasis. Wkly Epidemiol Rec 80 :202–212.
-
19
Helmy H, Fischer P, Farid HA, Bradley MH, Ramzy RM, 2004. Test strip detection of Wuchereria bancrofti amplified DNA in wild-caught Culex pipiens and estimation of infection rate by a PoolScreen algorithm. Trop Med Int Health 9 :158–163.
-
20
Taylor MJ, Bilo K, Cross HF, Archer JP, Underwood AP, 1999. 16S rDNA phylogeny and ultrastructural characterization of Wolbachia intracellular bacteria of the filarial nematode Brugia malayi, B. pahangi, and Wuchereria bancrofti. Exp Parasitol 91 :356–361.
-
21
Lulitanond V, Intapan PM, Pipitgool V, Choochote W, Maleewong W, 2004. Rapid detection of Wuchereria bancrofti in mosquitoes by LightCycler polymerase chain reaction and melting curve analysis. Parasitol Res 94 :337–341.
-
22
Farkas DH, Drevon AM, Kiechle FL, DiCarlo RG, Heath EM, Crisan D, 1996. Specimen stability for DNA-based diagnostic testing. Diagn Mol Pathol 5 :227–235.
-
23
Solomon AW, Holland MJ, Burton MJ, West SK, Alexander ND, Aguirre A, Massae PA, Mkocha H, Munoz B, Johnson GJ, Peeling RW, Bailey RL, Foster A, Mabey DC, 2003. Strategies for control of trachoma: observational study with quantitative PCR. Lancet 362 :198–204.
-
24
Lindahl T, 1993. Instability and decay of the primary structure of DNA. Nature 362 :709–715.
-
25
McGarry HF, Egerton GL, Taylor MJ, 2004. Population dynamics of Wolbachia bacterial endosymbionts in Brugia malayi. Mol Biochem Parasitol 135 :57–67.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 798 | 615 | 298 |
| Full Text Views | 257 | 5 | 0 |
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A REAL-TIME PCR-BASED ASSAY FOR DETECTION OF WUCHERERIA BANCROFTI DNA IN BLOOD AND MOSQUITOES
RAMAKRISHNA U. RAO
RAMAKRISHNA U. RAO
Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri; Research & Training Center on Vectors of Diseases, Ain Shams University, Cairo, Egypt; Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea; Department of Biological Sciences, Smith College, Northampton, Massachusetts
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LAURA J. ATKINSON
LAURA J. ATKINSON
Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri; Research & Training Center on Vectors of Diseases, Ain Shams University, Cairo, Egypt; Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea; Department of Biological Sciences, Smith College, Northampton, Massachusetts
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REDA M. R. RAMZY
REDA M. R. RAMZY
Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri; Research & Training Center on Vectors of Diseases, Ain Shams University, Cairo, Egypt; Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea; Department of Biological Sciences, Smith College, Northampton, Massachusetts
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HANAN HELMY
HANAN HELMY
Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri; Research & Training Center on Vectors of Diseases, Ain Shams University, Cairo, Egypt; Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea; Department of Biological Sciences, Smith College, Northampton, Massachusetts
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HODA A. FARID
HODA A. FARID
Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri; Research & Training Center on Vectors of Diseases, Ain Shams University, Cairo, Egypt; Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea; Department of Biological Sciences, Smith College, Northampton, Massachusetts
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MOSES. J. BOCKARIE
MOSES. J. BOCKARIE
Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri; Research & Training Center on Vectors of Diseases, Ain Shams University, Cairo, Egypt; Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea; Department of Biological Sciences, Smith College, Northampton, Massachusetts
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MELINDA SUSAPU
MELINDA SUSAPU
Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri; Research & Training Center on Vectors of Diseases, Ain Shams University, Cairo, Egypt; Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea; Department of Biological Sciences, Smith College, Northampton, Massachusetts
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SANDRA J. LANEY
SANDRA J. LANEY
Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri; Research & Training Center on Vectors of Diseases, Ain Shams University, Cairo, Egypt; Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea; Department of Biological Sciences, Smith College, Northampton, Massachusetts
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STEVEN A. WILLIAMS
STEVEN A. WILLIAMS
Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri; Research & Training Center on Vectors of Diseases, Ain Shams University, Cairo, Egypt; Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea; Department of Biological Sciences, Smith College, Northampton, Massachusetts
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GARY J. WEIL
GARY J. WEIL
Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri; Research & Training Center on Vectors of Diseases, Ain Shams University, Cairo, Egypt; Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea; Department of Biological Sciences, Smith College, Northampton, Massachusetts
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We developed and evaluated real-time polymerase chain reaction (PCR) assays for detecting Wuchereria bancrofti DNA in human blood and in mosquitoes. An assay based on detection of the W. bancrofti “LDR” repeat DNA sequence was more sensitive than an assay for Wolbachia 16S rDNA. The LDR-based assay was sensitive for detecting microfilarial DNA on dried membrane filters or on filter paper. We also compared real-time PCR with conventional PCR (C-PCR) for detecting W. bancrofti DNA in mosquito samples collected in endemic areas in Egypt and Papua New Guinea. Although the two methods had comparable sensitivity for detecting filarial DNA in reference samples, real-time PCR was more sensitive than C-PCR in practice with field samples. Other advantages of real-time PCR include its high-throughput capacity and decreased risk of cross-contamination between test samples. We believe that real-time PCR has great potential as a tool for monitoring progress in large-scale filariasis elimination programs.
Author Notes
- INTRODUCTION
- MATERIALS AND METHODS
- Detection of microfilaremia and filarial antigenemia.
- DNA isolation.
- Mosquito collection and DNA extraction.
- Real-time PCR assays for detection of W. bancrofti and Wolbachia DNA.
- C-PCR assay to detect Wuchereria bancrofti DNA in mosquitoes.
- Detection of W. bancrofti microfilariae in night blood samples.
- Ethical clearance.
- Data analysis.
- RESULTS
- Detection of W. bancrofti and Wolbachia DNA using real-time PCR.
- Specificity of W. bancrofti DNA detection by real-time PCR.
- Sensitivity of real-time PCR for detecting W. bancrofti DNA in dried blood from filariasis antigen card tests.
- Sensitivity of real-time PCR and C-PCR for detecting W. bancrofti DNA in Cx. pipiens and An. punctulatus mosquitoes.
- Effects of MDA on mosquito infection rates.
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Grove DI, Warren KS, Mahmoud AA, 1975. Algorithms in the diagnosis and management of exotic diseases. VI. The filariases. J Infect Dis 132 :340–352.
-
2
Weil GJ, Lammie PJ, Weiss N, 1997. The ICT filariasis test: a rapid-format antigen test for diagnosis of bancroftian filariasis. Parasitol Today 13 :401–404.
-
3
Lammie P, Weil G, Rahmah N, Kaliraj P, Steel C, Goodman D, Lakshmikanthan V, Ottesen E, 2004. Recombinant antigen based assays for the diagnosis and surveillance of lymphatic filariasis - a multicenter trial. Filaria J 3 :9.
-
4
Williams S, Laney S, Bierwert L, Saunders L, Boakye D, Fischer P, Goodman D, Helmy H, Hoti S, Vasuki V, Lammie P, Plichart C, Ramzy R, Ottesen E, 2002. Development and standardization of a rapid PCR-based method for the detection of Wuchereria bancrofti in mosquitoes for xenomonitoring the human prevalence of bancroftian filariasis. Ann Trop Med Parasitol 96 :S41–S46.
-
5
Ramzy R, Farid H, Kamal H, Ibrahim G, Morsy Z, Faris R, Weil G, 1997. A polymerase chain reaction-based assay for detection of Wuchereria bancrofti in human blood and Culex pipiens. Trans Roy Soc Trop Med Hyg 91 :156–160.
-
6
Zhong M, McCarthy J, Bierwert L, Lizotte-Waniewski M, Chanteau S, Nutman T, Ottesen E, Williams S, 1996. A polymerase chain reaction assay for detection of the parasite Wuchereria bancrofti in human blood samples. Am J Trop Med Hyg 54 :357–363.
-
7
McCarthy J, Zhong M, Gopinath R, Ottesen E, Williams S, Nutman T, 1996. Evaluation of a polymerase chain reaction-based assay for diagnosis of Wuchereria bancrofti infection. J Infect Dis 173 :1510–1514.
-
8
Fischer P, Liu X, Lizotte-Waniewski M, Kamal I, Ramzy R, Williams S, 1999. Development of a quantitative, competitive polymerase chain reaction-enzyme-linked immunosorbent assay for the detection of Wuchereria bancrofti DNA. Parasit Res 85 :176–183.
-
9
Abbasi I, Hamburger J, Githure J, Ochola J, Agure R, Koech D, Ramzy R, Gad A, Williams S, 1996. Detection of Wuchereria bancrofti DNA in patients sputum by the polymerase chain reaction. Trans Roy Soc Trop Med Hyg 90 :531–532.
-
10
Kluber S, Supali T, Williams SA, Liebau E, Fischer P, 2001. Rapid PCR-based detection of Brugia malayi DNA from blood spots by DNA detection test strips. Trans R Soc Trop Med Hyg 95 :169–170.
-
11
Bockarie MJ, Fischer P, Williams SA, Zimmerman PA, Griffin L, Alpers MP, Kazura JW, 2000. Application of a polymerase chain reaction-ELISA to detect Wuchereria bancrofti in pools of wild-caught Anopheles punctulatus in a filariasis control area in Papua New Guinea. Am J Trop Med Hyg 62 :363–367.
-
12
Chanteau S, Luquiaud P, Failloux AB, Williams SA, 1994. Detection of Wuchereria bancrofti larvae in pools of mosquitoes by the polymerase chain reaction. Trans R Soc Trop Med Hyg 88 :665–666.
-
13
Goodman DS, Orelus JN, Roberts JM, Lammie PJ, Streit TG, 2003. PCR and Mosquito dissection as tools to monitor filarial infection levels following mass treatment. Filaria J 2 :11.
-
14
Farid HA, Hammad RE, Hassan MM, Morsy ZS, Kamal IH, Weil GJ, Ramzy RM, 2001. Detection of Wuchereria bancrofti in mosquitoes by the polymerase chain reaction: a potentially useful tool for large-scale control programmes. Trans R Soc Trop Med Hyg 95 :29–32.
-
15
Vasuki V, Hoti SL, Sadanandane C, Jambulingam P, 2003. A simple and rapid DNA extraction method for the detection of Wuchereria bancrofti infection in the vector mosquito, Culex quinquefasciatus by Ssp I PCR assay. Acta Trop 86 :109–114.
-
16
Molyneux D, 2003. Lymphatic filariasis (elephantiasis) elimination: a public health success and development opportunity. Filaria J 2 :13.
-
17
Ottesen EA, 2002. Major progress toward eliminating lymphatic filariasis. N Engl J Med 347 :1885–1886.
-
18
World Health Organization, 2005. Global programme to eliminate lymphatic filariasis. Wkly Epidemiol Rec 80 :202–212.
-
19
Helmy H, Fischer P, Farid HA, Bradley MH, Ramzy RM, 2004. Test strip detection of Wuchereria bancrofti amplified DNA in wild-caught Culex pipiens and estimation of infection rate by a PoolScreen algorithm. Trop Med Int Health 9 :158–163.
-
20
Taylor MJ, Bilo K, Cross HF, Archer JP, Underwood AP, 1999. 16S rDNA phylogeny and ultrastructural characterization of Wolbachia intracellular bacteria of the filarial nematode Brugia malayi, B. pahangi, and Wuchereria bancrofti. Exp Parasitol 91 :356–361.
-
21
Lulitanond V, Intapan PM, Pipitgool V, Choochote W, Maleewong W, 2004. Rapid detection of Wuchereria bancrofti in mosquitoes by LightCycler polymerase chain reaction and melting curve analysis. Parasitol Res 94 :337–341.
-
22
Farkas DH, Drevon AM, Kiechle FL, DiCarlo RG, Heath EM, Crisan D, 1996. Specimen stability for DNA-based diagnostic testing. Diagn Mol Pathol 5 :227–235.
-
23
Solomon AW, Holland MJ, Burton MJ, West SK, Alexander ND, Aguirre A, Massae PA, Mkocha H, Munoz B, Johnson GJ, Peeling RW, Bailey RL, Foster A, Mabey DC, 2003. Strategies for control of trachoma: observational study with quantitative PCR. Lancet 362 :198–204.
-
24
Lindahl T, 1993. Instability and decay of the primary structure of DNA. Nature 362 :709–715.
-
25
McGarry HF, Egerton GL, Taylor MJ, 2004. Population dynamics of Wolbachia bacterial endosymbionts in Brugia malayi. Mol Biochem Parasitol 135 :57–67.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 798 | 615 | 298 |
| Full Text Views | 257 | 5 | 0 |
| PDF Downloads | 169 | 6 | 0 |