DETECTION AND DIFFERENTIATION OF FILARIAL PARASITES BY UNIVERSAL PRIMERS AND POLYMERASE CHAIN REACTION–RESTRICTION FRAGMENT LENGTH POLYMORPHISM ANALYSIS

SURANG NUCHPRAYOON Lymphatic Filariasis Research Unit, Department of Parasitology and Department of Pediatrics, Chula Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand; The W. Harry Feinstone Department of Molecular Microbiology and Immunology, The Johns Hopkins University, Bloomberg School of Hygiene and Public Health, Baltimore, Maryland

Search for other papers by SURANG NUCHPRAYOON in
Current site
Google Scholar
PubMed
Close
,
ALISA JUNPEE Lymphatic Filariasis Research Unit, Department of Parasitology and Department of Pediatrics, Chula Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand; The W. Harry Feinstone Department of Molecular Microbiology and Immunology, The Johns Hopkins University, Bloomberg School of Hygiene and Public Health, Baltimore, Maryland

Search for other papers by ALISA JUNPEE in
Current site
Google Scholar
PubMed
Close
,
YONG POOVORAWAN Lymphatic Filariasis Research Unit, Department of Parasitology and Department of Pediatrics, Chula Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand; The W. Harry Feinstone Department of Molecular Microbiology and Immunology, The Johns Hopkins University, Bloomberg School of Hygiene and Public Health, Baltimore, Maryland

Search for other papers by YONG POOVORAWAN in
Current site
Google Scholar
PubMed
Close
, and
ALAN L. SCOTT Lymphatic Filariasis Research Unit, Department of Parasitology and Department of Pediatrics, Chula Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand; The W. Harry Feinstone Department of Molecular Microbiology and Immunology, The Johns Hopkins University, Bloomberg School of Hygiene and Public Health, Baltimore, Maryland

Search for other papers by ALAN L. SCOTT in
Current site
Google Scholar
PubMed
Close
Restricted access

Filarial nematode parasites are a serious cause of morbidity in humans and animals. Identification of filarial infection using traditional morphologic criteria can be difficult and lead to misdiagnosis. We report on a polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP)–based method to detect and differentiate a broad range of filarial species in a single PCR. The first internal transcribed spacer 1 (ITS1) along with the flanking 18S and 5.8S ribosomal DNA (rDNA) were isolated and cloned from Wuchereria bancrofti, Brugia malayi, and Brugia pahangi. Sequence analysis identified conserved sites in the 18S and 5.8S rDNA sequence that could be used as universal priming sites to generate ITS1-distinctive PCR products that were useful for distinguishing filariae at the genus level. The addition of a digestion of the ITS1 PCR product with the restriction endonuclease Ase I generated a fragment profile that allowed differentiation down to the species level for W. bancrofti, B. malayi, B. pahangi, Dirofilaria immitis, and D. repens. The PCR-RFLP of ITS1 rDNA will be useful in diagnosing and differentiating filarial parasites in human, animal reservoir hosts, and mosquito vectors in disease-endemic areas.

Author Notes

  • 1

    Ottesen EA, Duke BO, Karam M, Behbehani K, 1997. Strategies and tools for the control/elimination of lymphatic filariasis. Bull World Health Organ 75 :491–503.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 2

    Molyneux DH, Bradley M, Hoerauf A, Kyelem D, Taylor MJ, 2003. Mass drug treatment for lymphatic filariasis and onchocerciasis. Trends Parasitol 19 :516–522.

  • 3

    World Health Organization, 1995. Bridging the Gaps. The World Health Report. Geneva: World Health Organization.

    • PubMed
    • Export Citation
  • 4

    Behbehani K, 1998. Candidate parasitic diseases. Bull World Health Organ 76 :S64–S67.

  • 5

    Guptavanij P, Harinasuta C, Sucharit S, Vutikes S, 1971. Studies on sub-periodic Brugia malayi in southern Thailand. Southeast Asian J Trop Med Public Health 2 :44–50.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 6

    Mak JW, 1987. Epidemiology of lymphatic filariasis. Ciba Found Symp 127 :5–14.

  • 7

    Zagaria N, Savioli L, 2002. Elimination of lymphatic filariasis: a public-health challenge. Ann Trop Med Parasitol 96 :S3–S13.

  • 8

    Dissanaike AS, 1979. Zoonotic aspects of filarial infections in man. Bull World Health Organ 57 :349–357.

  • 9

    Mak JW, Yen PK, Lim KC, Ramiah N, 1980. Zoonotic implications of cats and dogs in filarial transmission in peninsular Malaysia. Trop Geogr Med 32 :259–264.

  • 10

    Boreham PFL, 1988. Dirofilariasis in man. Boreham PFL, Atwell RB, eds. Dirofilariasis. Boca Raton, FL: CRC Press, 217–226.

    • PubMed
    • Export Citation
  • 11

    Muro A, Genchi C, Cordero M, Simon F, 1999. Human dirofilariasis in the European Union. Parasitol Today 15 :386–389.

  • 12

    Yoshimura H, Akao N, 1985. Current status of human dirofilariasis in Japan. Int J Zoonoses 12 :53–60.

  • 13

    Sukpanichnant S, Leenuttapong V, Dejsomritrutai W, Thakerngpol K, Wanachiwanawin W, Kachintorn U, Nitiyanant W, 1998. Pulmonary dirofilariasis in a patient with multisystem Langerhans cell histiocytosis: the first reported case in Thailand. J Med Assoc Thai 81 :722–727.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14

    Pampiglione S, Canestri Trotti G, Rivasi F, 1995. Human dirofilariasis due to Dirofilaria (Nochtiella) repens: a review of world literature. Parassitologia 37 :149–193.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 15

    Jariya P, Sucharit S, 1983. Dirofilaria repens from the eyelid of a woman in Thailand. Am J Trop Med Hyg 32 :1456–1457.

  • 16

    Lock JB, 1988. Dirofilaria sp.: taxonomic and distribution. Boreham PFL, Atwell RB, eds. Dirofilariasis. Boca Raton, FL: CRC Press, 1–28.

    • PubMed
    • Export Citation
  • 17

    Huynh T, Thean J, Maini R, 2001. Dipetalonema reconditum in the human eye. Br J Ophthalmol 85 :1391–1392.

  • 18

    Yen PK, Mak JW, 1978. Histochemical differentiation of Brugia, Wuchereria, Dirofilaria and Breinlia microfilariae. Ann Trop Med Parasitol 72 :157–162.

  • 19

    Nuchprayoon S, Sangprakarn S, Junpee A, Nithiuthai S, Chungpivat S, Poovorawan Y, 2003. Differentiation of Brugia malayi and Brugia pahangi by PCR-RFLP of ITS-1 and ITS-2. Southeast Asian J Trop Med Public Health 34 :67–73.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 20

    Nuchprayoon S, Yentakam S, Sangprakarn S, Junpee A, 2001. Endemic Bancroftian filariasis in Thailand: detection by Og4C3 antigen capture ELISA and the polymerase chain reaction. J Med Assoc Thai 84 :1300–1307.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 21

    Williams SA, Nicolas L, Lizotte-Waniewski M, Plichart C, Luquiaud P, Nguyen LN, Moulia-Pelat JP, 1996. A polymerase chain reaction assay for the detection of Wuchereria bancrofti in blood samples from French Polynesia. Trans R Soc Trop Med Hyg 90 :384–387.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 22

    Thanomsub BW, Chansiri K, Sarataphan N, Phantana S, 2000. Differential diagnosis of human lymphatic filariasis using PCR-RFLP. Mol Cell Probes 14 :41–46.

  • 23

    Triteeraprapab S, Kanjanopas K, Suwannadabba S, Sangprakarn S, Poovorawan Y, Scott AL, 2000. Transmission of the nocturnal periodic strain of Wuchereria bancrofti by Culex quinquefasciatus: establishing the potential for urban filariasis in Thailand. Epidemiol Infect 125 :207–212.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 24

    Triteeraprapab S, Karnjanopas K, Porksakorn C, Sai-Ngam A, Yentakam S, Loymak S, 2001. Lymphatic filariasis caused by Brugia malayi in an endemic area of Narathiwat Province, southern of Thailand. J Med Assoc Thai 84 :S182–S188.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 25

    Nuchprayoon S, Porksakorn C, Junpee A, Sanprasert V, Poovorawan Y, 2003. Comparative assessment of an Og4C3 ELISA and an ICT filariasis test: a study of Myanmar migrants in Thailand. Asian Pac J Allergy Immunol 21 :253–257.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 26

    World Health Organization, 1980. Staining and Identification. Manual for Basic Techniques for Health Laboratory. Geneva: World Health Organization, 209–214.

    • PubMed
    • Export Citation
  • 27

    Thompson JD, Higgins DG, Gibson TJ, 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22 :4673–4680.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 28

    McReynolds LA, DeSimone SM, Williams SA, 1986. Cloning and comparison of repeated DNA sequences from the human filarial parasite Brugia malayi and the animal parasite Brugia pahangi. Proc Natl Acad Sci U S A 83 :797–801.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 29

    Shah JS, Karam M, Piessens WF, Wirth DF, 1987. Characterization of an Onchocerca-specific DNA clone from Onchocerca volvulus. Am J Trop Med Hyg 37 :376–384.

  • 30

    Williams SA, DeSimone SM, McReynolds LA, 1988. Species-specific oligonucleotide probes for the identification of human filarial parasites. Mol Biochem Parasitol 28 :163–169.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 31

    Poole CB, Williams SA, 1990. A rapid DNA assay for the species-specific detection and quantification of Brugia in blood samples. Mol Biochem Parasitol 40 :129–136.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 32

    Siridewa K, Karunanayake EH, Chandrasekharan NV, Abeyewickreme W, Franzen L, Aslund L, Pettersson U, 1994. Cloning and characterization of a repetitive DNA sequence specific for Wuchereria bancrofti. Am J Trop Med Hyg 51 :495–500.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 33

    Lizotte MR, Supali T, Partono F, Williams SA, 1994. A polymerase chain reaction assay for the detection of Brugia malayi in blood. Am J Trop Med Hyg 51 :314–321.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 34

    Favia G, Lanfrancotti A, della Torre A, Cancrini G, Coluzzi M, 1997. Advances in the identification of Dirofilaria repens and Dirofilaria immitis by a PCR-based approach. Parassitologia 39 :401–402.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 35

    Chansiri K, Tejangkura T, Kwaosak P, Sarataphan N, Phantana S, Sukhumsirichart W, 2002. PCR based method for identification of zoonostic Brugia malayi microfilariae in domestic cats. Mol Cell Probes 16 :129–135.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 36

    Mar PH, Yang IC, Chang GN, Fei AC, 2002. Specific polymerase chain reaction for differential diagnosis of Dirofilaria immitis and Dipetalonema reconditum using primers derived from internal transcribed spacer region 2 (ITS2). Vet Parasitol 106 :243–252.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 37

    Gasser RB, 2001. Identification of parasitic nematodes and study of genetic variability using PCR approaches. Kennedy MW, Harnett W, eds. Parasitic Nematodes. Wallingford, United Kingdom: CAB International, 53–82.

    • PubMed
    • Export Citation
  • 38

    Wanji S, Tendongfor N, Esum M, Ndindeng S, Enyong P, 2003. Epidemiology of concomitant infections due to Loa loa, Mansonella perstans, and Onchocerca volvulus in rain forest villages of Cameroon. Med Microbiol Immunol (Berl) 192 :15–21.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 39

    Baneth G, Volansky Z, Anug Y, Favia G, Bain O, Goldstein RE, Harrus S, 2002. Dirofilaria repens infection in a dog: diagnosis and treatment with melarsomine and doramectin. Vet Parasitol 105 :173–178.

    • PubMed
    • Search Google Scholar
    • Export Citation
Past two years Past Year Past 30 Days
Abstract Views 140 110 13
Full Text Views 465 10 1
PDF Downloads 289 14 1
 
 
 
 
Affiliate Membership Banner
 
 
Research for Health Information Banner
 
 
CLOCKSS
 
 
 
Society Publishers Coalition Banner
Save