Molecular epidemiological surveillance of Africa and Asia imported malaria in Wuhan, Central China: comparison of diagnostic tools during 2011–2018

Abstract Background: Malaria remains a serious public health problem globally. As the elimination of indigenous malaria continues in China, imported malaria has gradually become a major health hazard. Well-timed and accurate diagnoses could support the timely implementation of therapeutic schedules, reveal the prevalence of imported malaria and avoid transmission of the disease.Methods: Blood samples were collected in Wuhan, China, from August 2011 to December 2018. All patients accepted microscopy and rapid diagnosis test (RDT) examinations. Subsequently, each of the positive or suspected positive cases was tested for four human-infectious Plasmodium species by using 18S rRNA-based nested PCR and Taqman probe-based real-time PCR. The results of the microscopy and the two molecular diagnostic methods were analysed. Importation origins were traced by country, and the prevalence of Plasmodium species was analysed by year.Results: A total of 296 blood samples, including 288 that were microscopy and RDT positive, 7 RDT and P. falciparum positive, and 1 suspected case, were collected and reanalysed. After application of the two molecular methods and sequencing, 291 cases including 245 P. falciparum, 15 P. vivax, 20 P. ovale, 6 P. malariae and 5 mixed infections (3 P. falciparum + P. ovale, 2 P. vivax + P. ovale) were confirmed. These patients had returned from Africa (95.53%) and Asia (4.47%). Although the prevalence displayed a small-scale fluctuation, the overall trend of the imported cases increased yearly. Conclusions: These results emphasize the necessity of combined utilization of the four tools for malaria diagnosis in clinic and in field surveys of potential risk regions worldwide including Wuhan.

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Molecular epidemiological surveillance of Africa and Asia imported malaria in Wuhan, Central China: comparison of diagnostic tools during 2011–2018

10.21203/rs.2.21390/v4 ◽

2020 ◽

Author(s):

Yiting Xie ◽

Kai Wu ◽

Weijia Cheng ◽

Tingting Jiang ◽

Yi Yao ◽

...

Keyword(s):

Plasmodium Species ◽

Public Health Problem ◽

Diagnostic Methods ◽

Imported Malaria ◽

Central China ◽

Molecular Diagnostic ◽

Diagnostic Tools ◽

Small Scale ◽

Suspected Case ◽

Blood Samples

Abstract Background Malaria remains a serious public health problem globally. As the elimination of indigenous malaria continues in China, imported malaria has gradually become a major health hazard. Well-timed and accurate diagnoses could support the timely implementation of therapeutic schedules, reveal the prevalence of imported malaria and avoid transmission of the disease. Methods Blood samples were collected in Wuhan, China, from August 2011 to December 2018. All patients accepted microscopy and rapid diagnosis test (RDT) examinations. Subsequently, each of the positive or suspected positive cases was tested for four human-infectious Plasmodium species by using 18S rRNA-based nested PCR and Taqman probe-based real-time PCR. The results of the microscopy and the two molecular diagnostic methods were analysed. Importation origins were traced by country, and the prevalence of Plasmodium species was analysed by year. Results A total of 296 blood samples, including 288 that were microscopy and RDT positive, 7 RDT and Plasmodium falciparum positive, and 1 suspected case, were collected and reanalysed. After application of the two molecular methods and sequencing, 291 cases including 245 P. falciparum, 15 Plasmodium vivax, 20 Plasmodium ovale, 6 Plasmodium malariae and 5 mixed infections (3 P. falciparum + P. ovale, 2 P. vivax + P. ovale) were confirmed. These patients had returned from Africa (95.53%) and Asia (4.47%). Although the prevalence displayed a small-scale fluctuation, the overall trend of the imported cases increased yearly. Conclusions These results emphasize the necessity of combined utilization of the four tools for malaria diagnosis in clinic and in field surveys of potential risk regions worldwide including Wuhan.

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Molecular epidemiological surveillance of Africa and Asia imported malaria in Wuhan, Central China: comparison of diagnostic tools during 2011–2018

10.21203/rs.2.21390/v1 ◽

2020 ◽

Author(s):

Yiting Xie ◽

Kai Wu ◽

Weijia Cheng ◽

Tingting Jiang ◽

Yi Yao ◽

...

Keyword(s):

Disease Transmission ◽

Plasmodium Species ◽

Public Health Problem ◽

Imported Malaria ◽

Central China ◽

Taqman Probe ◽

Epidemiological Surveillance ◽

Diagnostic Tools ◽

Small Scale ◽

Blood Samples

Abstract Background Malaria remains a serious public health problem globally. Along with indigenous malaria elimination in China, imported malaria gradually became a major hazard. Well-timed and accurate diagnosis could support immediate therapeutic schedule, reveal the prevalence of imported malaria and avoid the disease transmission. Method Blood samples were collected in Wuhan, China from August 2011 to December 2018. All patients first accepted microscopy and RDT examination. Subsequently, each of the positive or suspected positive cases was engaged in total four human Plasmodium species amplication by using 18S rRNA based nested PCR and Taqman probe based real-time PCR. Then performance of microscopy and two molecular diagnosis methods were analyzed. Importation origin was traced by country and prevalence of Plasmodium species was revealed by year. Results Total 296 blood samples containing 288 microscopy and RDT positive, 7 RDT P. falciparum positive and 1 suspected cases were collected and reanalyzed. After two molecular methods and sequencing detection, 291 cases including 245 P. falciparum , 15 P. vivax , 20 P. ovale , 6 P. malariae and 5 mixed infection (3 P. falciparum + P. ovale , 2 P. vivax + P. ovale ) were confirmed. These patients returned from Africa (95.53%) and Asia (4.47%). Although the prevalence displayed a small-scale fluctuation, the overall trend of the imported cases increased yearly. Conclusions Results emphasized the necessity of combined utilization of the four tools for malaria diagnosis in clinic and field survey around potential risk regions worldwide including Wuhan.

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Molecular epidemiological surveillance of Africa and Asia imported malaria in Wuhan, Central China: comparison of diagnostic tools during 2011–2018

10.21203/rs.2.20803/v1 ◽

2020 ◽

Author(s):

Yiting Xie ◽

Kai Wu ◽

Weijia Cheng ◽

Tingting Jiang ◽

Yi Yao ◽

...

Keyword(s):

Disease Transmission ◽

Plasmodium Species ◽

Public Health Problem ◽

Imported Malaria ◽

Central China ◽

Taqman Probe ◽

Epidemiological Surveillance ◽

Diagnostic Tools ◽

Small Scale ◽

Blood Samples

Abstract Background Malaria remains a serious public health problem globally. Along with indigenous malaria elimination in China, imported malaria gradually became a major hazard. Well-timed and accurate diagnosis could support immediate therapeutic schedule, reveal the prevalence of imported malaria and avoid the disease transmission. Method Blood samples were collected in Wuhan, China from August 2011 to December 2018. All patients first accepted microscopy and RDT examination. Subsequently, each of the positive or suspected positive cases was engaged in total four human Plasmodium species amplication by using 18S rRNA based nested PCR and Taqman probe based real-time PCR. Then performance of microscopy and two molecular diagnosis methods were analyzed. Importation origin was traced by country and prevalence of Plasmodium species was revealed by year. Results Total 296 blood samples containing 288 microscopy and RDT positive, 7 RDT P. falciparum positive and 1 suspected cases were collected and reanalyzed. After two molecular methods and sequencing detection, 291 cases including 245 P. falciparum , 15 P. vivax , 20 P. ovale , 6 P. malariae and 5 mixed infection (3 P. falciparum + P. ovale , 2 P. vivax + P. ovale ) were confirmed. These patients returned from Africa (95.53%) and Asia (4.47%). Although the prevalence displayed a small-scale fluctuation, the overall trend of the imported cases increased yearly. Conclusions Results emphasized the necessity of combined utilization of the four tools for malaria diagnosis in clinic and field survey around potential risk regions worldwide including Wuhan.

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LAMP in Neglected Tropical Diseases: A Focus on Parasites

Diagnostics ◽

10.3390/diagnostics11030521 ◽

2021 ◽

Vol 11 (3) ◽

pp. 521

Author(s):

Juan García-Bernalt Diego ◽

Pedro Fernández-Soto ◽

Antonio Muro

Keyword(s):

Neglected Tropical Diseases ◽

Treatment Success ◽

Public Health Problem ◽

Nucleic Acid Amplification ◽

Molecular Diagnostic ◽

Diagnostic Tools ◽

World Population ◽

Major Public Health Problem ◽

Tropical Diseases ◽

Early 21St Century

Neglected Tropical Diseases (NTDs), particularly those caused by parasites, remain a major Public Health problem in tropical and subtropical regions, with 10% of the world population being infected. Their management and control have been traditionally hampered, among other factors, by the difficulty to deploy rapid, specific, and affordable diagnostic tools in low resource settings. This is especially true for complex PCR-based methods. Isothermal nucleic acid amplification techniques, particularly loop-mediated isothermal amplification (LAMP), appeared in the early 21st century as an alternative to PCR, allowing for a much more affordable molecular diagnostic. Here, we present the status of LAMP assays development in parasite-caused NTDs. We address the progress made in different research applications of the technique: xenomonitoring, epidemiological studies, work in animal models and clinical application both for diagnosis and evaluation of treatment success. Finally, we try to shed a light on the improvements needed to achieve a true point-of-care test and the future perspectives in this field.

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The burden of congenital Chagas disease and implementation of molecular diagnostic tools in Latin America

BMJ Global Health ◽

10.1136/bmjgh-2018-001069 ◽

2018 ◽

Vol 3 (5) ◽

pp. e001069 ◽

Cited By ~ 21

Author(s):

Albert Picado ◽

Israel Cruz ◽

Maël Redard-Jacot ◽

Alejandro G Schijman ◽

Faustino Torrico ◽

...

Keyword(s):

Latin America ◽

Chagas Disease ◽

Diagnostic Algorithm ◽

Diagnostic Methods ◽

Diagnosis And Treatment ◽

Molecular Diagnostic ◽

Diagnostic Tools ◽

Molecular Tests ◽

Epidemiological Impact ◽

And Performance

It is estimated that between 8000 and 15 000 Trypanosoma cruzi infected babies are born every year to infected mothers in Chagas disease endemic countries. Currently, poor access to and performance of the current diagnostic algorithm, based on microscopy at birth and serology at 8–12 months after delivery, is one of the barriers to congenital Chagas disease (CCD) control. Detection of parasite DNA using molecular diagnostic tools could be an alternative or complement to current diagnostic methods, but its implementation in endemic regions remains limited. Prompt diagnosis and treatment of CCD cases would have a positive clinical and epidemiological impact. In this paper, we analysed the burden of CCD in Latin America, and the potential use of molecular tests to improve access to early diagnosis and treatment of T. cruzi infected newborns.

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Clinical Significance of Molecular Diagnostic Tools for Bacterial Bloodstream Infections: A Systematic Review

Interdisciplinary Perspectives on Infectious Diseases ◽

10.1155/2016/6412085 ◽

2016 ◽

Vol 2016 ◽

pp. 1-10 ◽

Cited By ~ 4

Author(s):

Jean Pierre Rutanga ◽

Therese Nyirahabimana

Keyword(s):

Systematic Review ◽

Antibiotic Therapy ◽

Clinical Significance ◽

Molecular Diagnosis ◽

Accurate Diagnosis ◽

Molecular Techniques ◽

Diagnostic Methods ◽

Diagnosis And Treatment ◽

Molecular Diagnostic ◽

Diagnostic Tools

Bacterial bloodstream infection (bBSI) represents any form of invasiveness of the blood circulatory system caused by bacteria and can lead to death among critically ill patients. Thus, there is a need for rapid and accurate diagnosis and treatment of patients with septicemia. So far, different molecular diagnostic tools have been developed. The majority of these tools focus on amplification based techniques such as polymerase chain reaction (PCR) which allows the detection of nucleic acids (both DNA and small RNAs) that are specific to bacterial species and sequencing or nucleic acid hybridization that allows the detection of bacteria in order to reduce delay of appropriate antibiotic therapy. However, there is still a need to improve sensitivity of most molecular techniques to enhance their accuracy and allow exact and on time antibiotic therapy treatment. In this regard, we conducted a systematic review of the existing studies conducted in molecular diagnosis of bBSIs, with the main aim of reporting on clinical significance and benefits of molecular diagnosis to patients. We searched both Google Scholar and PubMed. In total, eighteen reviewed papers indicate that shift from conventional diagnostic methods to molecular tools is needed and would lead to accurate diagnosis and treatment of bBSI.

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Development of a sensitive molecular diagnostic assay for detecting Borrelia burgdorferi DNA from the blood of Lyme disease patients by digital PCR

PLoS ONE ◽

10.1371/journal.pone.0235372 ◽

2020 ◽

Vol 15 (11) ◽

pp. e0235372

Author(s):

Srirupa Das ◽

Denise Hammond-McKibben ◽

Donna Guralski ◽

Sandra Lobo ◽

Paul N. Fiedler

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Digital Pcr ◽

Diagnostic Methods ◽

Molecular Diagnostic ◽

Diagnostic Assay ◽

Timely Manner ◽

Blood Samples ◽

Pcr Technology ◽

Analytical Processing

Lyme disease patients would greatly benefit from a timely, sensitive, and specific molecular diagnostic test that can detect the causal agent Borrelia burgdorferi at the onset of symptoms. Currently available diagnostic methods recommended by the Centers for Disease Control and Prevention for Lyme disease involve indirect serological tests that rely on the detection of a host-antibody response, which often takes more than three weeks to develop. With this process, many positive cases are not detected within a timely manner, preventing a complete cure. In this study, we have developed a digital polymerase chain reaction (PCR) assay that detects Lyme disease on clinical presentation with a sensitivity two-fold higher than that of the currently available diagnostic methods, using a cohort of patient samples collected from the Lyme disease endemic state of Connecticut, USA, in 2016–2018. Digital PCR technology was chosen as it is more advanced and sensitive than other PCR techniques in detecting rare targets. The analytical detection sensitivity of this diagnostic assay is approximately three genome copies of B. burgdorferi. The paucity of spirochetes in the bloodstream of Lyme disease patients has hindered the clinical adoption of PCR-based diagnostic tests. However, this drawback was overcome by using a comparatively larger sample volume, applying pre-analytical processing to the blood samples, and implementing a pre-amplification step to enrich for B. burgdorferi-specific gene targets before the patient samples are analyzed via digital PCR technology. Pre-analytical processing of blood samples from acute patients revealed that the best sample type for Lyme disease detection is platelet-rich plasma rather than whole blood. If detected in a timely manner, Lyme disease can be completely cured, thus limiting antibiotic overuse and associated morbidities.

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Diagnosis of Lymphatic Filariasis Using Immunochromatography and Microscopy in Three Local Government Areas of Cross River State, Nigeria

Nigerian Journal of Parasitology ◽

10.4314/njpar.v41i2.8 ◽

2020 ◽

Vol 41 (2) ◽

Author(s):

M Maurice ◽

E.U Sode ◽

I.B Otu-Bassey

Keyword(s):

Local Government ◽

Lymphatic Filariasis ◽

Neglected Tropical Diseases ◽

Age Groups ◽

Public Health Problem ◽

Diagnostic Methods ◽

Blood Samples ◽

Cross River ◽

Cross River State ◽

Significant Difference

Human Lymphatic filariasis (LF) caused by three types of filarial worms; Wuchereria bancrofti, Brugia malayi and Brugiatimori is one of the neglected tropical diseases and spread by bites of infected Anopheles, Culex, Aedes, Ochleratus andMansoni mosquitoes. The study evaluated diagnostic methods using blood samples from 420 consented participants in threeLocal Government Areas of Cross River State. Blood samples were examined following one hour of administration ofDiethylcarbamazine citrate for LF microfilaria microscopy (Knott’s concentration) and for LF antigen usingimmunochromatographic (ICT) method (Alere filariasis test strip). Of the 420 samples examined, 1.7% was found to bepositive using microscopy while 4.8% were positive using ICT. There was no significant difference in the diagnosis oflymphatic filariasis using microscopy and ICT among participants in the local government areas (χ2= 21.84, p>0.05). Of the214 males and 206 females examined, 4.2% males and 5.3% females tested positive using ICT while 1.4% males and 1.9%females were found positive using microscopy. The difference in the infection between gender was statistically significant(χ2=0.298, p<0.05). Participants aged 21-24 years had the highest prevalence of 19.4% while the least prevalence of 1.5%was observed among age group 9-12years using ICT. Also, no significant difference was observed in the diagnosis of LFamong the age groups (χ2= 19.88, p>0.05). The study showed that LF still remains a public health problem in Cross RiverState. Mass drug administration should be scaled up in the state so as to reduce and finally eradicate the disease. Keywords: Diagnosis, Lymphatic filariasis, Immunochromatography, Microscopy

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Development of a sensitive molecular diagnostic assay for detecting Borrelia burgdorferi DNA from blood of Lyme disease patients by digital PCR

10.1101/2020.06.16.154336 ◽

2020 ◽

Author(s):

Srirupa Das ◽

Denise Hammond-McKibben ◽

Donna Guralski ◽

Sandra Lobo ◽

Paul N. Fiedler

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Digital Pcr ◽

Diagnostic Methods ◽

Molecular Diagnostic ◽

Sample Type ◽

Diagnostic Assay ◽

Blood Samples ◽

Pcr Technology ◽

Analytical Processing

AbstractLyme disease patients would benefit greatly from a timely, sensitive and specific molecular diagnostic test that can detect the causal agent, Borrelia burgdorferi, at the onset of symptoms. Currently available diagnostic methods recommended by the Centers for Disease Control and Prevention for Lyme disease, involve indirect serological tests that rely on the detection of a host-antibody response which often takes more than three weeks to develop. This results in non-detection of many genuine cases on a timely basis, preventing complete cure. In this study we have developed a digital PCR (polymerase chain reaction) assay that detects Lyme disease on clinical presentation at twice the sensitivity of the currently available diagnostic methods, using a cohort of patient samples collected from the Lyme disease endemic state of Connecticut, USA in 2016-2018. Digital PCR technology was chosen as it is more advanced and sensitive than other PCR techniques in detecting rare targets and the lower limit of detection of this diagnostic assay was found to be three genome copies of B. burgdorferi. The paucity of spirochetes in the bloodstream of Lyme disease patients that hinders the clinical adoption of PCR-based diagnostic tests, was overcome by using a comparatively larger sample volume, pre-analytical processing of blood samples and a pre-amplification step to enrich for B. burgdorferi-specific gene targets before using the digital PCR technology to analyze patient samples. Pre-analytical processing of blood samples from acute patients revealed that the best sample type for Lyme disease detection is platelet-rich plasma and not whole blood. If detected on time, Lyme disease can be cured completely limiting the overuse of antibiotics and associated morbidities.

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