PCR-elisa for the detection of Brugia malayi infection using finger-prick blood

Rahmah, N.; A'shikin, A.Noor; Anuar, A. Khairul; Ariff, R.H.Tengku; Abdullah, Basima A.; Chan, Gloria; Williams, Steven A.

doi:10.1016/s0035-9203(98)91066-5

Cited by 22 publications

(7 citation statements)

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“…7,[9][10][11][12] It has been reported that a conventional PCR-ELISA without any internal control is a very sensitive tool for the detection of PCR products of the B. malayi Hha I repeat. 13 Our results confirmed this finding using a different labeling hapten and a different antibodyconjugate for the ELISA detection. In addition, we were able to show that an internal control can be used to avoid falsepositive results and the ELISA can be used to quantify PCR products to provide semiquantitative data.…”

Section: Discussionsupporting

confidence: 84%

“…4,5 Various sensitive polymerase chain reaction (PCR) assays have been established to detect W. bancrofti or B. malayi DNA in the human host or the mosquito vector. [6][7][8][9][10][11][12][13][14][15] Inhibition of a PCR, which may occur in both samples obtained from the mosquito vector and samples from the human host, is a common problem during DNA amplification and makes the standardization of PCR tests difficult. 16 Therefore, DNA constructs were used as internal standards (controls) in the PCR to avoid false-negative results in a diagnostic PCR.…”

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confidence: 99%

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Detection of DNA of nocturnally periodic Brugia malayi in night and day blood samples by a polymerase chain reaction-ELISA-based method using an internal control DNA.

Fischer

Supali²,

Wibowo³

et al. 2000

The American Journal of Tropical Medicine and Hygiene

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Abstract. An internal control was used in a polymerase chain reaction (PCR)-ELISA-based technique to detect the Hha I repeat of the filarial parasite Brugia malayi. A single microfilaria added to 200 l of blood was reliably detected. The assay was evaluated on field samples from persons living in an area endemic for Anopheles-transmitted, nocturnally periodic B. malayi in central Sulawesi, Indonesia. Examination of night blood of 138 individuals for the presence of microfilariae by filtration revealed 44 microfilaria carriers. All microfilaria carriers were also positive in the PCR-ELISA and, in addition, 14 more samples were proven to contain parasite DNA. The sensitivity of both methods was compared on night and on day blood samples collected from 113 persons. Whereas 37 microfilaria carriers were identified by filtration of night blood, no microfilariae were observed in the corresponding day blood samples. The PCR-ELISA result was positive in all 37 night blood samples of microfilaria carriers and in an additional 13 night blood samples without microfilariae. Parasite DNA was detected in 31 day blood samples of microfilaria carriers and in 3 day blood samples of amicrofilaremic persons. Assuming a sensitivity of the PCR-ELISA on night blood of 100%, the sensitivity of night blood filtration is 74% and that of the PCR-ELISA on day blood is 68%. These data suggest that the described PCR-ELISA method is capable of detecting infections with nocturnally periodic B. malayi in day blood samples. Therefore, this method may facilitate both the identification of endemic areas and the monitoring of control programs.Lymphatic filariasis, caused by Wuchereria bancrofti and Brugia malayi, has been targeted by the World Health Organization for elimination as a public health problem by the year 2020. 1 Therefore, control programs have been or will be launched world-wide with an emphasis on communitywide treatment with diethylcarbamazine (DEC) and ivermectin or albendazole.2 To monitor the success of such intervention programs, sensitive and cost effective diagnostic tools are required. It is estimated that 115 million people are infected with W. bancrofti and about 13 million with B. malayi. 3 In Asia, 1 of 5 filarial infections is due to B. malayi and in some countries this parasite is responsible for the majority of infections. 4,5

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Section: Discussionsupporting

confidence: 84%

mentioning

confidence: 99%

Detection of DNA of nocturnally periodic Brugia malayi in night and day blood samples by a polymerase chain reaction-ELISA-based method using an internal control DNA.

Fischer

Supali²,

Wibowo³

et al. 2000

The American Journal of Tropical Medicine and Hygiene

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“…The linear regression line from the correlation crossed the Y-axis at 47.31 HhaI copies/μl (SD = ±1.43) (Figure 1). The threshold of active infections was calculated to be 53.03 HhaI copies/μl (Y-intercept plus 4XSD) [28]. Transmigrant samples with HhaI copy numbers below the threshold were assumed to have DNA from L3 that were killed, but no active infection.…”

Section: Resultsmentioning

confidence: 99%

Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants

et al. 2014

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BackgroundLymphatic filariasis caused by Wuchereria bancrofti or Brugia spp. is a public health problem in developing countries. To monitor bancroftian filariasis infections, Circulating Filarial Antigen (CFA) test is commonly used, but for brugian infections only microfilariae (Mf) microscopy and indirect IgG4 antibody analyses are available. Improved diagnostics for detecting latent infections are required.MethodsAn optimized real-time PCR targeting the brugian HhaI repeat was validated with plasma from microfilariae negative Mongolian gerbils (jirds) infected with B. malayi. Plasma samples from microfilaremic patients infected with B. malayi or W. bancrofti were used as positive and negative controls, respectively. PCR results of plasma samples from a transmigrant population in a B. malayi endemic area were compared to those of life-long residents in the same endemic area; and to IgG4 serology results from the same population. To discriminate between active infections and larval exposure a threshold was determined by correlation and Receiver-Operating Characteristics (ROC) curve analyses.ResultsThe PCR detected HhaI in pre-patent (56 dpi) B. malayi infected jirds and B. malayi Mf-positive patients from Central Sulawesi, Indonesia. HhaI was also detected in 9/9 elephantiasis patients. In South Sulawesi 87.4% of the transmigrants and life-long residents (94% Mf-negative) were HhaI PCR positive. Based on ROC-curve analysis a threshold for active infections was set to >53 HhaI copies/μl (AUC: 0.854).ConclusionsThe results demonstrate that the HhaI PCR detects brugian infections with greater sensitivity than the IgG4 test, most notably in Mf-negative patients (i.e. pre-patent or latent infections).

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“…16 More recently, PCR using filarial ribosomal primers has been described as a valuable tool in confirming the parasite species. 17,18 These tests were not performed in our patient's biopsy due to unavailability of this technique at the time of the tissue referral, lack of need for further treatment due to localized disease, and failure to return for follow-up. The treatment depends on the clinical characteristics of each case.…”

Section: Discussionmentioning

confidence: 99%

A 32-Year-Old Immunocompetent Man With Submandibular Lymphadenopathy

Pérez¹,

Bush²,

Perdomo³

2006

Laboratory Medicine

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619Zoonotic filarial infections affecting humans have been reported worldwide, even in temperate countries. They might be caused by a variety of parasitic species, including Dirofilaria, Brugia, Onchocerca, Dipetalonema, Loaina, Meningonema, and certain Microfilariae. 1 While infection with Brugia is rather common in endemic areas of the world, it is a rare occurrence in the United States. In fact, less than 30 cases of locally acquired infection in human hosts have been described in the literature 2-7 since the first reported case in 1962. 8 Infection by Brugia species is considered a zoonotic disease transmitted by an insect vector, usually manifested by localized lymphadenitis or subcutaneous nodules in otherwise asymptomatic individuals when presenting in nonendemic geographic areas. The diagnosis is made by histopathologic demonstration of the parasite in the biopsied lesions. Since the majority of the reported cases showed localized disease, excisional biopsy was considered to be diagnostic and therapeutic simultaneously, making additional treatment unnecessary. We report the second case of zoonotic Brugia lymphadenitis acquired in Florida, and discuss the clinico-pathologic aspects of this unusual infectious disease. Case PresentationA 32-year-old immunocompetent man presented with submandibular lymphadenopathy after an episode of upper respiratory tract infection. He was treated at that time with a course of an oral cephalosporin for several weeks, with subsequent resolution of the respiratory symptoms and mild improvement of the lymphadenopathy. Of interest, he worked as an aquatic environmentalist in several southern states, including Florida. He had no history of traveling outside the United States.A computerized tomography (CT) scan of the head and neck disclosed a distinctly enlarged submandibular lymph node, completely separate from a radiologically unremarkable submandibular salivary gland. He underwent surgical excision of the enlarged lymph node. Clinical ProgressionHistopathological examination revealed reactive follicular lymphoid hyperplasia and a focus of granulomatous inflammation, elicited by a parasite located in the subcapsular sinus (Images 1 and 2). The tissue sections and paraffin block were forwarded to the Division of Infectious and Parasitic Diseases at the Armed Forces Institute of Pathology (AFIP) in Washington, DC, for further identification and speciation of the parasite. The organism was described by the consultant pathologists as an adult female nematode possessing a thin cuticle with thickened lateral ridges and paired uteri containing unfertilized eggs. The size and morphological features of the parasite were consistent with Brugia filarial species [a confirmatory polymerase chain reaction (PCR) test using ribosomal primers was not performed due to unavailability of this test at the time of consultation]. Laboratory findings including complete blood cell count with white cell differential and a basic metabolic profile showed values within normal reference ranges. There wa...

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PCR-elisa for the detection of Brugia malayi infection using finger-prick blood

Cited by 22 publications

References 6 publications

Detection of DNA of nocturnally periodic Brugia malayi in night and day blood samples by a polymerase chain reaction-ELISA-based method using an internal control DNA.

Detection of DNA of nocturnally periodic Brugia malayi in night and day blood samples by a polymerase chain reaction-ELISA-based method using an internal control DNA.

Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants

A 32-Year-Old Immunocompetent Man With Submandibular Lymphadenopathy

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