Nested PCR detection of Plasmodium malariae from microscopy confirmed P. falciparum samples in endemic area of NE India

Abstract

Malaria is a complex and evolved infectious human disease that is endemic in north-eastern states of india, where malaria control relies on early active and passive case detection and prompt treatment through malaria camps and village level health workers (Dhiman et al. 2011). since malaria associated with fever and other symptoms is the most common diagnosis in rural settings, the incorrect diagnosis may have severe public health implications (Anash et al. 2010). A considerable proportion of patients treated for malaria actually do not have malaria (Prasad 2009). Furthermore, missed out true malaria cases may act as epicentres for disease transmission at a local level. resistance to anti-malarials, decay in health infrastructure, population movement and socio-political unrest are contributing factors to the spread of malaria (gardner et al. 2002). Accurate diagnosis of malaria cases could be useful in lowering such conjectures and ultimately reducing malaria burden in endemic settings.Until recently, microscopic examination of blood smears has been a standard method for malaria diagnosis by convenience (Anash et al. 2010). Microscopy, however, requires well trained technical staff and it is also rather labour intensive and time consuming (Maltha et al. 2010). Hand-held non-microscopic immunochromatographic rapid diagnostic tests (rDt's) are simple, sensitive and specific, and good results have been achieved with them in various endemic regions. since their introduction during the late 1990's, rDt's have undergone many changes and currently three-band rDt's for detecting both Plasmodium falciparum Welch, 1897 specific antigen and Plasmodium-specific antigen (PAN-specific) have been developed (Moody 2002). the performance of the rDt's in the diagnosis of malaria has been validated throughout the malaria endemic regions (Moody 2002, Anash et al. 2010, however, the rDt's were unable to identify mixed infections, owing to the common PAN-specific antibody capture band, when compared to the polymerase chain reaction (Pcr) assay. in addition, the rDt's have not been found suitable to diagnose malaria where parasite counts were as low as < 50 parasite/µl (iqbal et al. 1999).the plDH based optiMAl-it ® has some limitations, but as on today, it has been proved to be one of the best rDt available for on spot detection of malaria parasite Nested PCR detection of Plasmodium malariae from microscopy confirmed P. falciparum samples in endemic area of NE India Abstract: the present study evaluates the performance of optiMAl-it ® test and nested Pcr assay in detection of malaria parasites. A total of 76 randomly selected blood samples collected from two malaria endemic areas were tested for malaria parasites using microscopy and optiMAl-it ® test in the field. Pcr assays were performed in the laboratory using DNA extracted from blood spots of the same samples collected on the FtA ™ classic cards. of the total of 61 field confirmed malaria positive samples, only 58 (95%) were detected positive using microscopy in the laboratory....

Plasmodium malariae in the Colombian Amazon region: you don’t diagnose what you don’t suspect

“…[38, 39, 57, 58], together with 98–100% estimations for detecting P. vivax [38, 59, 60]. Similarly, PCR estimated higher prevalence values for the species evaluated and for certain types of co-infection, such increases having been observed in previous studies for both simple and mixed infections [22, 23, 27, 47, 48]. This result highlights PCR’s potential for confirming a clinical suspicion of malaria, in spite of being expensive and not available in health centres having limited resources [61].…”

“…The corresponding samples were in turn diagnosed by TBS as negative or simple infection caused by either P. vivax or P. falciparum , the predominant and regular species in the target region. Lack of quartan malaria detection by microscopy may have been related to TBS limitations per se as P. malariae is characterized by sustaining low infection rates and low parasitaemia [47, 48]. Similarly, the common loss of cells’ distinctive characteristics in samples treated for TBS can also account for overlooking P. malariae infection, given that it hampers accurate species identification [15, 22, 23, 27, 48].…”

Genetic polymorphism and amino acid sequence variation in Plasmodium falciparum GLURP R2 repeat region in Assam, India, at an interval of five years

“…Northeastern states of India report high P. falciparum cases and malaria-attributed deaths annually [ 9 , 10 ]. The entire northeast region has unique geography conducive to malaria spread and also its poor socio-economic conditions increase transmission risk [ 11 – 13 ].…”

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

“…However, the use of microscopic examination has been restricted because it requires well-trained laboratory personnel and shows less sensitive at low parasitaemia [22,23,24]. Actually, different observers may make two to three fold discrepancies in parasite quantification [25]. More importantly, mixed infections of different Plasmodium species were frequently missed [26].…”