IJCRR

IJCRR - 9(8), April, 2017

Pages: 36-40

Date of Publication: 01-May-2017

Microbial Forensics- Issues and Challenges

Author: Moumita Sinha, I. Arjun Rao

Category: Healthcare

Abstract:Bioterrorism has been known to exist since hundreds of years by utilising microorganisms as weapons. The terrorist attack using anthrax in the fall of 2001 in U.S. highlighted this kind of utilisation of microorganisms. To criminally prosecute with the help of forensic evidence the perpetrator is necessary and such attacks of bioterrorism are very few. A comprehensive technological network is necessary to strengthen defence against bio crimes by acquiring knowledge of various fields needs to be developed. One such new field connected microbiology and scientific science is known as Microbial Forensics. It utilizes advanced molecular methods like DNA microarray examination and DNA fingerprinting and so on to relate the wellspring of the causative specialist with a particular individual or gathering by measuring varieties between related strains. Excellent affirmation and quality control models for microbial legal sciences will guarantee very dependable outcomes that will stand up in the official courtroom. The more exact and refined a microbial framework turns into, the more appropriate rules for examinations will be characterized. An incorporated approach towards building up this field of microbial crime scene investigation should be taken after, to meet the difficulties of bioterrorism all the more successfully.

Keywords: Microbial, Bioterrorism, Forensics, DNA, Fingerprinting

Full Text:

INTRODUCTION

The human body is a place of residence for trillions of microorganisms, living both outwardly and additionally characteristically. Such microorganisms are massively differing and extend from microbes, infection, parasites, to protozoa. The viroids are additionally thought to be a piece of it, while a few researchers consider them as non-living organisms (1). The passing extraneous gathering of organisms is gotten from the outer environment, while the inborn microorganisms are available in the human body for a more drawn out period of time. The review in connection to such microorganisms is alluded to as microbiology.
The microbial legal sciences that have been assigned as a specialized train connecting microbiology and legal pharmaceutical perpetrated to the examination of bioterrorism and bio-wrongdoing. The branch grasps a colossal chance of legal science pummelled with microbiology, which includes examination of microorganisms or their poisons and additionally the constituents used to figure, stockpile and dispersal of the pathogenic organism. The mammoth noticeable quality of this novel branch is imperative for the contemporary space attributable to the reality the information and mastery controlled by the felonious terrorist could be disastrous to the humankind. The impact and heading of such an assault have been seen complex extends ever. In any case, it was genuinely reflected when the United States watched a risk of a Bacillus anthracis assault in the United states in the year 2001. Detecting the capability of such science, there was another branch planned in crime scene investigation, alluded to “Microbial Forensics” (2).
On the other hand, the essential belief system to use forensic science with the end goal of the criminal examination started in the year 1995. It was clearly helpful to utilize forensic solution as opposed to the normal examination inferable from the view that examinations with the guide of forensic science could be easily performed without the acquire capacity of witnesses, opportune reportage, files available for evaluation, and unanticipated confirmation. In those days it was seen that one day a minor number of research experts would flourish in such an activity, by having associate with microbial crime scene investigation, utilizing safe keeping, and utilizing tricky assets all through dissemination of a natural operator (3).

The Bio weapons concept

Microorganisms are potential weapons since they can be obtained from a single creature or cells and not at all like nuclear weapons, can be delivered in a little foundation with low capital speculation and without the requirement for modern instruments and skilled labourers (4). In spite of the fact that there are numerous techniques to make biological weapons the basic alternative is discharge microorganism that is possibly pathogenic or might be lessened to more pathogenic structures and afterward discharged into the group which makes it most deadly than some other weapon. Any microbiological specialist can be effectively utilized as weaponry, of which the essential concentration will be an operator which is more environment safe and pathogenic. With the utilization of genetic engineering, even an ecological commensal might be outfitted with a compelling poison creating quality. This would make it naturally steady and a very powerful weapon. A rundown of the conceivable species as a bio danger is past the extent of composing as any living being might be lessened and utilized as the trigger. For a rundown of high ready living beings, per users are alluded to Friedrich et.al (5) and the Centre for Disease Control (CDC) site (6). The recognition strategy must incorporate a convention that can be delicate, particular and fast. Molecular finding favours the most extreme in such a setting. It’s relevance in a creating nation can however be testing, attributable to assets. Additionally numerous life forms required in the crime might be absolutely obscure or the DNA succession in part known which makes it hard to build up a molecular assay.

Objectives of Forensic Microbiologists

The fundamental objectives of microbial crime scene investigation is distinguish and organize natural dangers, recognize the powerless populace, make a data database and create conventions for ID which incorporates deciding special hereditary marks, protein marks, create programs for guaranteeing the legitimacy of results and continually overhaul in view of existing writing (7, 8). Recognizing and organizing the objective is more troublesome than normally suspected. The life form that should be organized might be absolutely obscure or the life form may not be a human pathogen, yet rather a plant pathogen which can incur high financial harm (9, 10). Recognizable proof of helpless populace would be similarly testing. Making a data database would help in directing towards the outcomes, however unexpectedly it can likewise help the criminal in recognizing those living beings which are not ordered and hence clear a path for the expansion of a living being more hard to distinguish. Legitimacy of utilization of such a database ought to be clear for similar reasons. The database that is utilized will include contribution from different established field’s comprehensive of microbiology, genomics, scientific strategies, science and immaculatescience (11). A potential rundown of dangers ought to be promptly accessible to all concerned. This ought to be implying as need targets.
Distinguishing proof of powerless populace is similarly testing. Making a data database would help in directing towards the outcomes, however unexpectedly it can likewise help hoodlums in distinguishing those life forms which are not sorted and in this manner clear a path for the multiplication of a living being harder to recognize. Authentic utilization of such a database ought to be clear for similar reasons. The database that is utilized ought to include contributions from different established field’s comprehensive of microbiology, genomics, forensic methods, chemistry and pure science.

Control of Quality and Substantiation

The quality confirmation and quality control program is an unavoidable part of any research and microbial legal sciences are also imparted. Building up a convention for identification may make utilization of routine demonstrative approaches, for which quality rules exist. Furthermore, methods that have not experienced approval may likewise must be utilized, particularly when the living being is obscure or once in a while experienced which is right now not prescribed by any directing documentations. Such outcomes may not be exceptionally dependable but rather can make signs for judging the conceivable living being. This is of exceptional worry as the life form being managed may not be much known to the logical world. Scientific Working Group on Microbial Genetics and Forensics (SWGMGF) sets up and supports rules as well as norms for quality frameworks distinguishing procedures and techniques, characterize criteria for learning frameworks and above all fill in as an accomplished asset on issues as they emerge (8). The SWGMGF characterizes the rules and overhauls it as and when required. The advancement of these rules helps the research centre to perform different measurable investigations and host the outcomes as legitimate and consistent with the best of logical learning accessible to that day. Extra stringent guidelines are required (11) contrasted with routine reviews as the issues include lawful matters and information will be depended upon vigorously. An incorrect quality administration or absence of value control may misdirect the last conclusion. The chain of authority ought to be immaculate
to get flawless outcomes, particularly the biological confirmations acquired in this specific situation. A standard operating procedure (SOP) may not be accessible dependably and regularly direction from different organizations might be required and suppositions considered. Additionally, developing an approval plan and its execution will help the cause (11). The improvement of guidelines ought to be founded on benchmarks of human DNA writing, clinical research facilities norms and International Standards Organization.

Laboratory based approach

Laboratories involved in analysis of forensic microbiological evidences must be geared up to deal with chain-of custody documentation, assured storage of facts, tracking of individual items of evidence and their derivatives and all the legal requirements for handling evidence. Chain- of-custody customizes the records in uninterrupted chain of records screening who had contend with the evidence, where and under which conditions [temperature, time etc.] the material had been stored and whether access to the samples was restricted knowledge available to that day. Further rigorous set of laws are required (12) compared to tedious appraisals as the matters involve official issues and records will be relied upon heavily. A flawed quality management or lack of quality control may delude the ultimate finale. The chain of custody should be impeccable to obtain perfect results, especially the biologic evidences obtained in this context. A standard operating procedure (SOP) may not be available always and often guidance from other institutions may be required and opinions considered. In addition, constructing a validation plan and its execution will help the cause (13). The development of rulesshould be based on standards of human DNA typing, clinical laboratories standards and International Standards Organization (ISO).

The knowledge of microbial forensics becomes an integral factor when examiners are given a presumed case with a strange introduction or in a place where the dispersion of illness is irregular. If there should be an occurrence of a biocrime, commonly, the laboratory that acquires the specimen as the standard test is the one to first raise a doubt. In the event that a strongly doubted is summoned to be imparted to an investigative body or national reference hub, particularly when a strain that looks genetically built or test examination demonstrates multi-strains of conceivable aetiology (14).

The steps required for the examination are basically the same as the examination of a characteristic flare-up. Be that as it may, they are more requesting than the routine analytic or epidemiological test (15). The example accumulation is of most extreme significance. The specimens to be gathered incorporate each material found in the scene which is marked with time and site of accumulation. The name of the individual who has gathered the specimen ought to likewise be specified. The code of practice ought to be the same regardless of the sort of the example from, a group or person. Microbiological proof could incorporate; suitable examples of the microbial operators, protein poisons, nucleic acids, clinical examples from casualties, research centre hardware, spread gadgets and their substance, natural specimens, sullied garments, or follow confirm particular to the procedure that delivered as well as weaponries the organic specialist. On the criminological front, the technique for gathering ought to be touchy, dependable and powerful to secure the nearness of conceivable life form (16). Convenient natural examining is of tremendous value as it might be quickly devastated and the confirmation of deliberate spread might be lost (8). Every specimen ought to be considered possibly unsafe and prepared just in an all around prepared research facility, or in a perfect world sent to a reference lab outfitted with stringent biosecurity levels (15).

Techniques used in elucidating the causative agent(s)

The recognizable proof of microbial specialists – as characterized by the SIBCA handbook - can be temporary [presumptive], when immunological strategies, nucleic corrosive recognition or development and metabolic examines have been tried positive. Distinguishing proof is affirmed by the blend of no less than two of the previously mentioned criteria. Unambiguous recognizable proof requires development and in vivo thinks about [animal models] that demonstrate the pathogenicity of the specialist. In any case, creature models ought to be stayed away from for moral reasons at whatever point conceivable. Organic specialists can be hard to develop because of test defilement, low number of microbes or pre-treatment of patients with anti-microbial. A few microscopic organisms are fussy [F. tularensis, Brucella spp.] and require extraordinary supplement media, and some need delayed development times [Brucella spp.] (17)

Phenotypical attributes, for example, anti-toxin helplessness and biochemical response profiles, weakness to particular phages, settlement morphology and others are not generally dependable. Transformations of operators can be initiated or designed, however normally happening atypical strains have likewise been discovered e.g. among Bacillus anthracis and Yersinia pestis confines which can bring about misidentification and treatment disappointment (18). Commercial biochemical distinguishing proof frameworks are not streamlined for these operators and can bring about misidentification. Numerous antimicrobial resistances can happen through the regular flat quality exchange or by hereditary. Common imperviousness to a huge number of antimicrobials is regular for Burkholderia pseudo mallei.Francisella tularensis is normally impervious to penicillins and cephalosporines. An extremely hazardous multidrug safe strain of Yersinia pestis has been segregated from a patient with bubonic torment in Madagascar. This strain conveys a self-transmissible plasmid with a hereditary spine likewise pervasive among Escherichia coli, Klebsiella spp. furthermore, Salmonella spp. giving abnormal state imperviousness to streptomycin, tetracyclin, chloramphenicol, and sulfonamides (19). These actualities underline the significance of development and the appraisal of antimicrobial weakness notwithstanding more fast analytic devices. A polyphasic approach for recognizable proof and writing will maintain a strategic distance from issues because of atypical genotype and phenotype, restraint, or absence of specificity or affectability of measures. Treatment of select operators is profoundly unsafe and bulky and confined to labs with bio safety-level 3 regulation. Biosafety-level 3 research facilicustody documentation, assured storage of facts, tracking of individual items of evidence and their derivatives and all the legal requirements for handling evidence. Chain- of-custody customizes the records in uninterrupted chain of recordsscreening who had contend with the evidence, where and under which conditions [temperature, time etc.] the material had been stored and whether access to the samples was restricted knowledge available to that day. Further rigorous set of laws are required (12) compared to tedious appraisals as the matters involve official issues and records will be relied upon heavily. A flawed quality management or lack of quality control may delude the ultimate finale. The chain of custody should be impeccable to obtain perfect results, especially the biologic evidences obtained in this context. A standard operating procedure (SOP) may not be available always and oftenguidance from other institutions may be required and opinionsconsidered. In addition, constructing a validation plan and its execution will help the cause (13). The development of rules should be based on standards of human DNA typing, clinical laboratories standards and International Standards Organization (ISO). The knowledge of microbial forensics becomes an integral factor when examiners are given a presumed case with a strange introduction or in a place where the dispersion of illness is irregular. If there should be an occurrence of a biocrime, commonly, the laboratory that acquires the specimen as the standard test is the one to first raise a doubt. In the event that a strongly doubted is summoned to be imparted to
an investigative body or national reference hub, particularly when a strain that looks genetically built or test examination demonstrates multi-strains of conceivable aetiology (14). The steps required for the examination are basically the same as the examination of a characteristic flare-up. Be that as it may, they are more requesting than the routine analytic or epidemiological test (15). The example accumulation is of most extreme significance. The specimens to be gathered incorporate each material found in the scene which is marked with time and site of accumulation. The name of the individual who has gathered the specimen ought to likewise
be specified. The code of practice ought to be the same regardless of the sort of the example from, a group or person. Microbiological proof could incorporate; suitable examples
of the microbial operators, protein poisons, nucleic acids, clinical examples from casualties, research centre hardware, spread gadgets and their substance, natural specimens, sullied
garments, or follow confirm particular to the procedure that delivered as well as weaponries the organic specialist. On the criminological front, the technique for gathering ought to be touchy, dependable and powerful to secure the nearness of conceivable life form (16). Convenient natural examining is of tremendous value as it might be quickly devastated and the confirmation of deliberate spread might be lost (8). Every specimen ought to be considered possibly unsafe and prepared just in an all around prepared research facility, or in a perfect world sent to a reference lab outfitted with stringent biosecurity levels (15).

Techniques used in elucidating the causative agent(s)

The recognizable proof of microbial specialists – as characterized by the SIBCA handbook - can be temporary [presumptive], when immunological strategies, nucleic corrosive recognition or development and metabolic examines have been tried positive. Distinguishing proof is affirmed by the blend of no less than two of the previously mentioned criteria. Unambiguous recognizable proof requires development and in vivo thinks about [animal models] that demonstrate the pathogenicity of the specialist. In any case, creature models ought to be stayed away from for moral reasons at whatever point conceivable. Organic specialists can be hard to develop because of test defilement, low number of microbes or pre-treatment of patients with anti-microbial. A few microscopic organisms are fussy [F. tularensis, Brucella spp.] and require extraordinary supplement media, and some need delayed development times [Brucella spp.] (17) Phenotypical attributes, for example, anti-toxin helplessness and biochemical response profiles, weakness to particular phages, settlement morphology and others are not generally dependable. Transformations of operators can be initiated or designed, however normally happening atypical strains have likewise been discovered e.g. among Bacillus anthracis and Yersinia pestis confines which can bring about misidentification and treatment disappointment (18). Commercial biochemical distinguishing proof frameworks are not streamlined for these operators and can bring about misidentification. Numerous antimicrobial resistances can happen through the regular flat quality exchange or by hereditary control. Common imperviousness to a huge number of antimicrobials is regular for Burkholderia pseudo mallei. Francisella tularensis is normally impervious to penicillins and cephalosporines. An extremely hazardous multidrug safe strain of Yersinia pestis has been segregated from a patient with bubonic torment in Madagascar. This strain conveys a self-transmissible plasmid with a hereditary spine likewise pervasive among Escherichia coli, Klebsiella spp. furthermore, Salmonella spp. giving abnormal state imperviousness to streptomycin, tetracyclin, chloramphenicol, and sulfonamides (19). These actualities underline the significance of development and the appraisal of antimicrobialweakness notwithstanding more fast analytic devices. A polyphasic approach for recognizable proof and writing will maintain a strategic distance from issues because of atypical genotype and phenotype, restraint, or absence of specificity or affectability of measures. Treatment of select operators is profoundly unsafe and bulky and confined to labs with bio safety-level 3 regulation. Biosafety-level 3 research facilities must be worked by uncommon controls that require e.g. a modern ventilation framework and individual defensive equipment [e.g. FFP3 masks, overalls, face shields, gloves etc.].

1. Nucleic Acid Amplification Techniques

Several PCR assays are very particular and delicate and abbreviate the time required to build up a determination in correlation with ordinary PCR conventions, development, and biochemical distinguishing proof techniques. Thusly, continuous PCR measures have been produced for the distinguishing proof of Bacillus anthracis, Brucella spp., Burkholderia
mallei and Burkholderia pseudomallei, Francisella tularensisand Yersinia pestis (20). PCR results can be false negative because of deficient nature of clinical specimens, low number of microscopic organisms in tests, DNA degradation, inhibitory substances and incorrect DNA readiness.

2. Serology

Seroconversion may demonstrate the presentation to a particular operator before. Be that as it may, seroconversion can be normal simply following a few days or weeks and is of little use for quickly diagnosing contaminations brought on by exceptionally pathogenic specialists. It will be hard to arrange serological examinations [including follow-up tests] when a fear monger assault causes mass losses that need medicinal treatment or when the circumstance is convoluted by disasters and civil wars in the meantime. Different immunological measures have likewise been utilized to distinguish pathogens in tests of patients and natural examples. Manual test units can be utilized as bed-side tests and are helpful under field conditions, however, clinical approvals barely exist and most tests are “for logical utilize as it were”. Immunochromatographic sidelong stream tests have been created e.g. for brucellosis, tularemia, and plague (21-24). Confinements of these immunological examines are that they are habitually not accessible economically, not sufficiently particular, or have not been approved and authorized for use in people or creatures. Also, cross-responses may bring about false positives and adjusted or miss antigenic structures can bring about false negatives.

Typing and strain identification

Differences among microorganisms must be evaluated to figure out if strains are from a similar source or ancestry or from an alternate beginning. The exactness and accuracy will rely on upon the writing strategy, expected transformation rates, and different attributes of the creature. In court, researchers may need to measure the unwavering quality of a relationship among strains decided to utilize molecular phylogenetic examinations. This will build up the likelihood of relationship to a specific wellspring of disease (25). Methods for forensic microbiology can be fundamentally the same as those being utilized for phylogenetic and epidemiological examinations e.g. for food-borne outbreaks. Molecular epidemiological instruments utilized for genotyping are most encouraging and have been connected in the
past to explain the birthplace of organic specialists. Particularly entire genome sequencing and bioinformatic apparatuses for examination of genomes are intense instruments, however,
many-sided quality and expenses are still restrictive for routine application. In a few sections of the exceedingly recommendable book “Microbial Forensics” by Bruce Budowle and numerous other “authors” of this new logical train it was exhibited that exclusive exceptionally particular learning of microbial hereditary qualities will permit an appraisal of the significance of writing results acquired by Multi-locus Sequence Typing [MLST], Variable Number of Tandem Repeats
[VNTR], Single Nucleotide Polymorphisms [SNPs] examination or other writing instruments (26). Approval of writing examines and information of vast accumulations of strains from everywhere throughout the world are critical for microbial legal examinations. Writing techniques ought to be reproducible, stable amid the review time frame, pertinent to each confine, segregating among detaches, and separation ought to be concordant with the epidemiological picture (27). DNA sequence-based information is strong, versatile, simple to contrast and amiable with electronic examination for phylogeographical and epidemiological reviews. Be that as it may, the nature of open get to arrangement databases relies on upon the exactness of submitted successions and is thus infrequently not firm (26).

Indian scenario

To the best of our insight, there is no reported instance of natural fighting in India. This might be translated as no case happening or as the absence of criminological microbiology work up. Remembering the last probability, microbial crime scene investigation has an unfathomable potential in India. As a nation which is regularly undermined by fear assaults, there are most likely natural weapons will be made utilization of by different psychological militant outfits sooner rather than later. Foundation of a national association which coordinates ability of pros from different fields of science will turn out to be helpful regardless of the cost that will be caused in making and keeping up such a group. Three parts will be significant to set up a completely useful National Microbial Forensic Laboratory. The first would be a learning focus made out of databases on genomics, microbiology, legal strategies, SOPs, prove examines, for example, fingerprinting, bioinformatics and institutionalized devices. The second segment will be upkeep of solid organizations between the current government, the research centre in charges, researchers and exploring operators. The third segment will be quality control and approval of more up to date examines (28).

CONCLUSION

Microbial forensics is a logical train devoted to breaking down confirmation from a bioterrorism demonstration, biocrime or incidental microorganism/poison discharge for attribution purposes. The unlawful utilization of natural specialists postures generous threats to people, general wellbeing, the earth, the economies of countries, and worldwide peace. A national and universal cooperative approach can be utilized to deal with the hazard of bioterrorism by setting up a national and global reference research centre in this manner guaranteeing straightforwardness of investigation and strict activity against all bio wrongdoing culprits. Considering all the logical realities as of now examined, “Microbial Forensics” ought to be a perfect prerequisite in India.

ACKNOWLEDGEMENT

Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.

References:

1. Lwoff A. “The concept of virus”. J. Gen. Microbiol.1956; 17 (2); 239–253.
2. Budowle B, Schutzer SE, Einseln A, Kelley LC, Walsh AC, et al. Public health. Building microbial forensics as a response to bioterrorism. Science 2003; 301: 1852–1853.
3. Marshall JC; Gastrointestinal flora and its alterations in critical illness. Curr Opin Clin Nutr Metab Care, 1999; 2: 405–411.
4. Priyabrata Pattnaik, Krisnamurthy Sekhar. Forensics for tracing microbial signatures: Biodefense perspective and preparedness for the unforeseen. Indian Jn of Biotech 2008; 7:23-31.
5. Friedrich Frischknecht, The history of biological warfare, Human experimentation, modern nightmares and lone mad men in the twentieth century; EMBO reports 2003;Vol 4 Special issue.
6. http://www.bt.cdc.gov/agent/agentlist.asp. Accessed on January 2nd, 2016.
7. Microbial forensics: A new forensic discipline, Sharad Jain, Ashish Kumar, Pratima Gupta, Ramjee Prasad, Dept. of Microbiology, Himalayan Institute of Medical Sciences, Dehradun,
JIAFM, 2005; 27 (2).
8. Defining a New Forensic Discipline: Microbal Forensics Bruce Budowle Laboratory Division, Federal Bureau of Investigation, Washington, DC, U.S.A. March 2003. www.promega.com. Accessed on Dec 20, 2011
9. Whitby S, Rogers P. Anticrop biological warfare—Implications of the Iraqi and US programmes. Defense Analysis 1997;13: 303–318
10. Whitby SM. The potential use of plant pathogens against crops. Microbes Infect 2001; 3(1):73–80
11. Budowle, Quality sample collection, handling, and preservation for an effective microbial forensics program. Appl. Environ. Microbiol.; 72: 6431-6438.
12. Chakrakodi Narayana Varun, Kuruvilla Thomas S, Furtado Zevita. (2012). Microbial Forensics- Past, Present and Future. International Journal of Biological & Medical Research. Int J Biol Med Res. 2012; 3(2): 1546-1549.
13. Kaur M, Gupte S, Aggarwal P, Manhas A, Bala M ,Mahajan S. (2014). Methods in Microbial Forensics. J Punjab Acad Forensic Med Toxicol ;14(1).
14. Treadwell TA, Koo D, Kuker K and Khan AS. (2003). Epidemiologic clues to bioterrorism. Public Health Rep; 118:92–98.
15. Ronald M. (2004). Atlas. Microbial Forensics- Taking Diagnostic Microbiology to the Next Level. Clinical Microbiology Newsletter. 26, 13
16. National science and technology council (2015) National strategy to support research in microbial forensics attribution investigations and national security.http://www.whitehouse.gov/files/ documents/ostp/NSTC%20 Reports/National%20MicroForensics% 20R&DStrategy%202009%20UNLIMITED%20DISTRIBUTION. pdf. Accessed on Nov 24.
17. Wilmoth BA, Chu MC and Quan TJ (1996) Identification of Yersinia pestis by BBL Crystal Enteric/Nonfermenter Identification System. J Clin Microbiol. (34), 2829-30.
18. Welch TJ, Fricke WF, Mc Dermott P.F,White DG, Rosso ML, Rasko DA et al (2007) Multiple antimicrobial resistance in plague: an emerging public health risk. PLoS One. (2),e309 EOF
19. Jones SW, Dobson ME, Francesconi SC, Schoske R and Crawford R (2005) DNA assays for detection, identification, and individualization of select agent microorganisms. Croat Med J. (46), 522-9.
20. Chanteau S, Rahalison L, Ralafiarisoa L, Foulon J, Ratsitorahina M, Ratsifasoamanana L et al (2003) Development and testing of a rapid diagnostic test for bubonic and pneumonic plague. Lancet. (361), 211-6
21. Mizanbayeva S, Smits HL, Zhalilova K, Abdoel TH, Kozakov S aand Ospanov KS (2009) The evaluation of a user-friendly lateral flow assay for the serodiagnosis of human brucellosis in Kazakhstan. Diagn Microbiol Infect Dis. (65), 14-20.
22. Splettstoesser W, Guglielmo-Viret V, Seibold E and Thullier (2010) Evaluation of an immunochromatographic test for rapid and reliable serodiagnosis of human tularemia and detection of Francisella tularensis-specific antibodies in sera from different mammalian species. J Clin Microbiol. (48), 1629-34
23. Tomaso H, Thullier P, Seibold E, Guglielmo V, Buckendahl A, Rahalison L et al (2007) Comparison of hand-held test kits, immunofluorescence microscopy, enzyme-linked immunosorbent assay, and flow cytometric analysis for rapid presumptive identification of Yersinia pestis. J Clin Microbiol, (45):3404-7.
24. Gonzalez-Candelas F, Bracho M.A, Moya A (2003) Molecular epidemiology and forensic genetics: application to a hepatitis C virus transmission event at a hemodialysis unit. J Infect Dis. (187), 352-8.
25. Van Belkum A, Tassios PT, Dijkshoorn L, Haeggman S, Cookson B, Fry N.K et al (2007) Guidelines for the validation and application of typing methods for use in bacterial epidemiology. Clin Microbiol. (3), 1-46.
26. Tomaso H and Neubauyer H (2011) Forensic Microbiology. In: Vieira D.N (ed), Forensic Medicine-From old problems to new challenges, Intech.
27. Bruce Budowle (2008) Criteria for validation of methods in microbial forensics, Applied And Environmental Microbiology. Vol. 74, No. 18, P. 5599–5607.
28. Bhatia Mohit, Mishra Bibhabati, Thakur Archana, Dogra Vinita, Loomba Poonam Sood (2016) Concept of Forensic Microbiology and its Applications. SMU Medical Journal. Volume – 3, No. 1.v